Method for treating tumors

Abstract

Provided are compositions for use in treating a subject having a small cell lung cancer (SCLC) - derived tumor having a high tumor mutational burden (TMB) status.SOLUTION: Provided are methods of treating a subject having an SCLC-derived tumor having a TMB status, comprising administering to the subject a monotherapy comprising an anti-PD-1 antibody or a combination therapy comprising an anti-PD-1 antibody and an anti-CTLA-4 antibody. Also provided are methods of identifying a subject suitable for treatment with an anti-PD-1 antibody or a combination therapy comprising an anti-PD-1 antibody and an anti-CTLA-4 antibody, comprising determining the TMB status in a biological sample of the subject. A high TMB status identifies the patient as suitable for treatment with an anti-PD-1 antibody or antigen-binding portion thereof. TMB status can be determined by sequencing nucleic acids in the tumor and identifying genomic alterations, e.g., somatic non-synonymous mutations, in the sequenced nucleic acids.SELECTED DRAWING: None

Description

[Technical Field]

[0001] FIELD OF THE INVENTION The present invention provides methods of treating a subject having a tumor with a high Tumor Mutational Burden (TMB) status, e.g., SCLC, comprising administering to the subject an anti-PD-1 antibody alone ("monotherapy") or an anti-PD-1 antibody in combination with an anti-CTLA-4 antibody. [Background technology]

[0002] Background of the Invention Human cancers possess numerous genetic and epigenetic alterations, producing neoantigens that may be recognizable by the immune system (Sjoblom et al., Science (2006) 314(5797):268-274). The adaptive immune system, consisting of T and B lymphocytes, has potent anticancer potential, with broad capacity and exquisite specificity to respond to a wide variety of tumor antigens. Furthermore, the immune system exhibits considerable plasticity and memory components. Successfully utilizing all these properties of the adaptive immune system makes immunotherapy unique among all cancer treatment modalities.

[0003] Until recently, cancer immunotherapy has focused considerable effort on methods to enhance antitumor immune responses through adoptive transfer of activated effector cells, immunization against relevant antigens, or nonspecific immunostimulators such as cytokines. However, over the past decade, intensive efforts to develop specific immune checkpoint pathway inhibitors have begun to offer novel immunotherapeutic approaches for cancer treatment, including the development of antibodies such as nivolumab and pembrolizumab (formerly lambrolizumab; USAN Council Statement, 2013), which specifically bind to the programmed cell death-1 (PD-1) receptor and block the inhibitory PD-1 / PD-1 ligand pathway (Topalian et al., 2012a, b; Topalian et al., 2014; Hamid et al., 2013; Hamid and Carvajal, 2013; McDermott and Atkins, 2013).

[0004] PD-1 is an important immune checkpoint receptor expressed by activated T cells and B cells and mediates immunosuppression. PD-1 is a member of the CD28 family of receptors, which also includes CD28, CTLA-4, ICOS, PD-1, and BTLA. Two cell surface glycoprotein ligands for PD-1, programmed cell death ligand-1 (PD-L1) and programmed cell death ligand-2 (PD-L2), have been identified and are expressed on antigen-presenting cells and numerous human cancers. Binding to PD-1 has been shown to downregulate both T cell activation and cytokine secretion. Inhibition of the PD-1 / PD-L1 interaction mediates potent antitumor activity in preclinical models (U.S. Patents 8,008,449 and 7,943,743), and the use of antibody inhibitors of the PD-1 / PD-L1 interaction for cancer treatment has entered clinical trials (Brahmer et al., 2010; Topalian et al., 2012a; Topalian et al., 2014; Hamid et al., 2013; Brahmer et al., 2012; Flies et al., 2011; Pardoll, 2012; Hamid and Carvajal, 2013).

[0005] Nivolumab (formerly known as 5C4, BMS-936558, MDX-1106, or ONO-4538) is a fully human IgG4(S228P) PD-1 immune checkpoint inhibitor antibody that selectively blocks interaction with PD-1 ligands (PD-L1 and PD-L2), thereby blocking downregulation of antitumor T cell function (US Pat. No. 8,008,449; Wang et al., 2014). Nivolumab has demonstrated activity in a variety of advanced solid tumors, including renal cell carcinoma (renal adenocarcinoma or Grawitz tumor), melanoma, and non-small cell lung cancer (NSCLC) (Topalian et al., 2012a; Topalian et al., 2014; Drake et al., 2013; WO 2013 / 173223). Summary of the Invention [Problem to be solved by the invention]

[0006] The immune system and its response to immunotherapy are complex. Furthermore, anti-cancer drugs can vary in effectiveness based on unique patient characteristics. Thus, there is a need for targeted therapeutic strategies that identify patients who are likely to respond to specific anti-cancer drugs, thereby improving clinical outcomes for patients diagnosed with cancer. [Means for solving the problem]

[0007] Summary of the Invention The present invention provides a method for treating a subject with a tumor derived from small cell lung cancer (SCLC), comprising administering to the subject a therapeutically effective amount of an antibody or antigen-binding portion thereof that specifically binds to the programmed death-1 (PD-1) receptor and inhibits PD-1 activity (an "anti-PD-1 antibody"), wherein the tumor has a high tumor mutational burden (TMB) status. The present invention also provides a method for treating a subject with a tumor derived from SCLC, comprising administering to the subject a therapeutically effective amount of an anti-PD-1 antibody and an antibody or antigen-binding portion thereof that specifically binds to CTLA-4 (an "anti-CTLA-4 antibody"), wherein the tumor has a high TMB status. In certain embodiments, the method further comprises measuring the TMB status of a biological sample obtained from the subject.

[0008] The present invention also provides a method for identifying a subject having an SCLC-derived tumor and suitable for treatment with an anti-PD-1 antibody, the method comprising measuring the TMB status of a biological sample from the subject, wherein the TMB status is high TMB. In certain embodiments, the method further comprises administering an anti-PD-1 antibody to the subject. In certain embodiments, the method further comprises administering an anti-PD-1 antibody and an anti-CTLA-4 antibody to the subject.

[0009] The present invention also provides a method for identifying a subject having an SCLC-derived tumor and suitable for combination therapy comprising an anti-PD-1 antibody and an anti-CTLA-4 antibody, the method comprising measuring the TMB status of a biological sample from the subject, wherein the TMB status is high. In certain embodiments, the method further comprises administering to the subject an anti-PD-1 antibody and an anti-CTLA-4 antibody.

[0010] In some embodiments, TMB status is determined by sequencing nucleic acid in tumor and identifying genomic alterations in the sequenced nucleic acid.In some embodiments, genomic alterations include one or more somatic mutations.In some embodiments, genomic alterations include one or more non-synonymous mutations.In certain embodiments, genomic alterations include one or more missense mutations.In other specific embodiments, genomic alterations include one or more alterations selected from the group consisting of base pair substitution, base pair insertion, base pair deletion, copy number alteration (CNA), gene rearrangement, and any combination thereof.

[0011] In certain embodiments, TMB status is determined by genome sequencing, exome sequencing and / or genome profiling.In some embodiments, genome profile comprises at least 300 genes, at least 305 genes, at least 310 genes, at least 315 genes, at least 320 genes, at least 325 genes, at least 330 genes, at least 335 genes, at least 340 genes, at least 345 genes, at least 350 genes, at least 355 genes, at least 360 genes, at least 365 genes, at least 370 genes, at least 375 genes, at least 380 genes, at least 385 genes, at least 390 genes, at least 395 genes or at least 400 genes.In certain embodiments, genome profile comprises at least 325 genes.

[0012] ABL1, BRAF, CHEK1, FANCC, GATA3, JAK2, MITF, PD CD1LG2, RBM10, STAT4, ABL2, BRCA1, CHEK2, FANCD2, GATA4, JAK3, MLH1, PDGF RA, RET, STK11, ACVR1B, BRCA2, CIC, FANCE, GATA6, JUN, MPL, PDGFRB, RICTO R, SUFU, AKT1, BRD4, CREBBP, FANCF, GID4(C17orf39), KAT6A(MYST3), MRE11 A, PDK1, RNF43, SYK, AKT2, BRIP1, CRKL, FANCG, GLI1, KDM5A, MSH2, PIK3C2B ROS1, TAF1, AKT3, BTG1, CRLF2, FANCL, GNA11, KDM5C, MSH6, PIK3CA, RPTOR. TBX3, ALK, BTK, CSF1R, FAS, GNA13, KDM6A, MTOR, PIK3CB, RUNX1, TERC, AMER 1(FAM123B), C11orf30(EMSY), CTCF, FAT1, GNAQ, KDR, MUTYH, PIK3CG, RUNX1 T1, TERT(Type), APC, CARD11, CTNNA1, FBXW7, GNAS, KEAP1, MYC, PIK3R 1, SDHA, TET2, AR, CBFB, CTNNB1, FGF10, GPR124, KEL, MYCL(MYCL1), PIK3R2. SDHB, TGFBR2, ARAF, CBL, CUL3, FGF14, GRIN2A, KIT, MYCN, PLCG2, SDHC, and TNF AIP3, ARFRP1, CCND1, CYLD, FGF19, GRM3, KLHL6, MYD88, PMS2, SDHD, TNFRSF1 4. ARID1A, CCND2, DAXX, FGF23, GSK3B, KMT2A(MLL), NF1, POLD1, SETD2, TOP 1, ARID1B, CCND3, DDR2, FGF3, H3F3A, KMT2C(MLL3), NF2, POLE, SF3B1, and TOP2A ARID2, CCNE1, DICER1, FGF4, HGF, KMT2D(MLL2), NFE2L2, PPP2R1A, SLIT2, T P53, ASXL1, CD274, DNMT3A, FGF6, HNF1A, KRAS, NFKBIA, PRDM1, SMAD2, TSC1.ATM, CD79A, DOT1L, FGFR1, HRAS, LMO1, NKX2-1, PREX2, SMAD3, TSC2, ATR, CD79B, EGFR, FGFR2, HSD3B1, LRP1B, NOTCH1, PRKAR1A, SMAD4, TSHR , ATRX, CDC73, EP300, FGFR3, HSP90AA1, LYN, NOTCH2, PRKCI, SMARCA4, U2AF1, AURKA, CDH1, EPHA3, FGFR4, IDH1, LZTR1, NOTCH3, PRKDC, SMARC B1, VEGFA, AURKB, CDK12, EPHA5, FH, IDH2, MAGI2, NPM1, PRSS8, SMO, VHL, AXIN1, CDK4, EPHA7, FLCN, IGF1R, MAP2K1, NRAS, PTCH1, SNCAIP, WI SP3, AXL, CDK6, EPHB1, FLT1, IGF2, MAP2K2, NSD1, PTEN, SOCS1, WT1, BAP1, CDK8, ERBB2, FLT3, IKBKE, MAP2K4, NTRK1, PTPN11, SOX10, XPO1, BA RD1, CDKN1A, ERBB3, FLT4, IKZF1, MAP3K1, NTRK2, QKI, SOX2, ZBTB2, BCL2, CDKN1B, ERBB4, FOXL2, IL7R, MCL1, NTRK3, RAC1, SOX9, ZNF217, BC L2L1, CDKN2A, ERG, FOXP1, INHBA, MDM2, NUP93, RAD50, SPEN, ZNF703, BCL2L2, CDKN2B, ERRFI1, FRS2, INPP4B, MDM4, PAK3, RAD51, SPOP, BCL6, The gene may comprise one or more genes selected from the group consisting of CDKN2C, ESR1, FUBP1, IRF2, MED12, PALB2, RAF1, SPTA1, BCOR, CEBPA, EZH2, GABRA6, IRF4, MEF2B, PARK2, RANBP2, SRC, BCORL1, CHD2, FAM46C, GATA1, IRS2, MEN1, PAX5, RARA, STAG2, BLM, CHD4, FANCA, GATA2, JAK1, MET, PBRM1, RB1, STAT3, and any combination thereof.

[0013] In certain embodiments, the method further comprises identifying genomic alterations in one or more of ETV4, TMPRSS2, ETV5, BCR, ETV1, ETV6, and MYB.

[0014] In some embodiments, high TMB is at least 210, at least 215, at least 220, at least 225, at least 230, at least 235, at least 240, at least 245, at least 250, at least 255, at least 260, at least 265, at least 270, at least 275, at least 280, at least 285, at least 290, at least 295, at least 300, at least 305, at least 310, at least 315, at least 320, at least 325, at least 330, at least 335, at least 340, at least 345, at least 350, at least 360, at least 365, at least 370, at least 375, at least 380, at least 385, at least 390, at least 400, at least 410, at least 420, at least 430, at least 440, at least 450, at least 460, at least 470, at least 480, at least 490, at least 500, at least 510, at least 520, at least 530, at least 540, at least 550, at least 560, at least 570, at least 580, at least 590, at least 600, at least 610, at least 620, at least 630, at least 640, at least 650, at least 660, at least 670, at least 680, at least 690, at least 700, at least 710, at least 720, at least 730, at least 740, at least 750 In some embodiments, the marker has a score of at least 355, at least 360, at least 365, at least 370, at least 375, at least 380, at least 385, at least 390, at least 395, at least 400, at least 405, at least 410, at least 415, at least 420, at least 425, at least 430, at least 435, at least 440, at least 445, at least 450, at least 455, at least 460, at least 465, at least 470, at least 475, at least 480, at least 485, at least 490, at least 495, or at least 500. In other embodiments, high TMB has a score of at least 215, at least 220, at least 221, at least 222, at least 223, at least 224, at least 225, at least 226, at least 227, at least 228, at least 229, at least 230, at least 231, at least 232, at least 233, at least 234, at least 235, at least 236, at least 237, at least 238, at least 239, at least 240, at least 241, at least 242, at least 243, at least 244, at least 245, at least 246, at least 247, at least 248, at least 249, or at least 250. In certain embodiments, high TMB has a score of at least 243.

[0015] In some embodiments, the method further comprises comparing the subject's TMB status with a control TMB value. In some embodiments, the subject's TMB status is within the highest quantile of the control TMB value. In other embodiments, the subject's TMB status is within the highest tertile of the control TMB value.

[0016] In some embodiments, biological sample is tumor tissue biopsy, for example, formalin-fixed, paraffin-embedded tumor tissue or fresh-frozen tumor tissue.In other embodiments, biological sample is liquid biopsy.In some embodiments, biological sample comprises one or more of blood, serum, plasma, exoRNA, circulating tumor cell, ctDNA and cfDNA.

[0017] In certain embodiments, the subject has a tumor with a high neoantigen load. In other embodiments, the subject's T cell repertoire is expanded.

[0018] In some embodiments, the SCLC comprises small cell carcinoma. In some embodiments, the SCLC comprises mixed small cell carcinoma. In some embodiments, the SCLC is recurrent SCLC.

[0019] In some embodiments, the subject has received at least one, at least two, at least three, at least four, or at least five previous therapies for treating tumors. In some embodiments, the previous therapies include chemotherapy. In some embodiments, the chemotherapy includes platinum-based therapy. In some embodiments, the platinum-based therapy includes a platinum-based anti-neoplastic agent selected from the group consisting of cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenanthriplatin, picoplatin, satraplatin, and any combination thereof. In some embodiments, the platinum-based therapy includes cisplatin.

[0020] In some embodiments, the anti-PD-1 antibody cross-competes with nivolumab for binding to human PD-1. In other embodiments, the anti-PD-1 antibody binds to the same epitope as nivolumab. In some embodiments, the anti-PD-1 antibody is a chimeric antibody, a humanized antibody, a human monoclonal antibody, or an antigen-binding portion thereof. In other embodiments, the anti-PD-1 antibody comprises a heavy chain constant region of a human IgG1 isotype or a human IgG4 isotype. In certain embodiments, the anti-PD-1 antibody is nivolumab or pembrolizumab.

[0021] In some embodiments, the anti-PD-1 antibody is administered at a dose ranging from 0.1 mg / kg to 10.0 mg / kg body weight every 2, 3, or 4 weeks. In some embodiments, the anti-PD-1 antibody is administered at a dose of 5 mg / kg or 10 mg / kg body weight every 3 weeks. In other embodiments, the anti-PD-1 antibody is administered at a dose of 5 mg / kg body weight every 3 weeks. In yet other embodiments, the anti-PD-1 antibody is administered at a dose of 3 mg / kg body weight every 2 weeks.

[0022] In some embodiments, the anti-PD-1 antibody is administered as a fixed dose. In some embodiments, the anti-PD-1 antibody is administered as a fixed dose of at least about 200 mg, at least about 220 mg, at least about 240 mg, at least about 260 mg, at least about 280 mg, at least about 300 mg, at least about 320 mg, at least about 340 mg, at least about 360 mg, at least about 380 mg, at least about 400 mg, at least about 420 mg, at least about 440 mg, at least about 460 mg, at least about 480 mg, at least about 500 mg, or at least about 550 mg. In other embodiments, the anti-PD-1 antibody is administered as a fixed dose about every 1 week, 2 weeks, 3 weeks, or 4 weeks.

[0023] In some embodiments, the anti-CTLA-4 antibody is a chimeric, humanized or human monoclonal antibody or a portion thereof. In some embodiments, the anti-CTLA-4 antibody comprises a heavy chain constant region of human IgG1 isotype. In some embodiments, the anti-CTLA-4 antibody is ipilimumab. In some embodiments, the anti-CTLA-4 antibody is tremelimumab. In some embodiments, the anti-CTLA-4 antibody cross-competes with ipilimumab for binding to human CTLA-4.

[0024] In some embodiments, the anti-CTLA-4 antibody is administered at a dose ranging from at least about 0.1 mg / kg to at least about 10.0 mg / kg body weight about every 1, 2, 3, or 4 weeks. In some embodiments, the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg or about 3 mg / kg body weight. In some embodiments, the anti-CTLA-4 antibody is administered at a fixed dose. In some embodiments, the anti-CTLA-4 antibody is administered about every 2 weeks. In some embodiments, the anti-CTLA-4 antibody is administered about every 3 weeks.

[0025] In one embodiment, the anti-PD-1 antibody is administered at a dose of about 3 mg / kg body weight about every three weeks and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg body weight about every three weeks. In one embodiment, the anti-PD-1 antibody is administered at a dose of about 1 mg / kg body weight about every three weeks and the anti-CTLA-4 antibody is administered at a dose of about 3 mg / kg body weight about every three weeks.

[0026] In one embodiment, (i) the anti-PD-1 antibody is administered at a dose of about 1 mg / kg body weight and the anti-CTLA-4 antibody is administered at a dose of about 3 mg / kg body weight about every 3 weeks for 4 cycles, and then (ii) the anti-PD-1 antibody is administered at a dose of about 3 mg / kg body weight about every 2 weeks.

[0027] In certain embodiments, the subject exhibits progression-free survival for at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 1 year, at least about 18 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years after administration.

[0028] In other embodiments, the subject exhibits overall survival for at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 1 year, at least about 18 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years after administration.

[0029] In still other embodiments, subjects exhibit an objective response rate of at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100%.

[0030] Other features and advantages of the present disclosure will be apparent from the following detailed description and examples, which should not be construed as limiting. The contents of all references cited throughout this specification, including scientific articles, press reports, GenBank entries, patents, and patent applications, are expressly incorporated herein by reference.

[0031] Implementation E1. A method of treating a subject having a tumor derived from small cell lung cancer (SCLC), comprising administering to the subject a therapeutically effective amount of an antibody or antigen-binding portion thereof (an "anti-PD-1 antibody") that specifically binds to the programmed cell death-1 (PD-1) receptor and inhibits PD-1 activity, wherein the tumor has a high tumor mutational burden (TMB) status.

[0032] E2. A method of treating a subject having a tumor derived from SCLC, comprising administering to the subject therapeutically effective amounts of an anti-PD-1 antibody and an antibody or antigen-binding portion thereof that specifically binds to CTLA-4 ("anti-CTLA-4 antibody"), wherein the tumor has a TMB-high status.

[0033] E3. The method of E1 or E2, further comprising determining the TMB status of a biological sample obtained from the subject.

[0034] E4. A method for identifying a subject having an SCLC-derived tumor and suitable for treatment with an anti-PD-1 antibody, comprising measuring the TMB status of a biological sample from the subject, wherein the TMB status is high TMB.

[0035] E5. A method for identifying a subject having an SCLC-derived tumor and suitable for combination therapy comprising an anti-PD-1 antibody and an anti-CTLA-4 antibody, the method comprising measuring the TMB status of a biological sample from the subject, wherein the TMB status is high TMB.

[0036] E6. The method of E4, further comprising administering to the subject an anti-PD-1 antibody.

[0037] E7. The method of E5, further comprising administering to the subject an anti-PD-1 antibody and an anti-CTLA-4 antibody.

[0038] E8. The method of any of E1-E7, wherein TMB status is determined by sequencing nucleic acid in the tumor and identifying genomic alterations in the sequenced nucleic acid.

[0039] E9. The method of E8, wherein the genomic alterations include one or more somatic mutations.

[0040] E10. The method of E8 or E9, wherein the genomic alteration comprises one or more non-synonymous mutations.

[0041] E11. The method according to any one of E8 to E10, wherein the genomic alteration comprises one or more missense mutations.

[0042] E12. The method described in any of E8 to E11, wherein the genomic alteration comprises one or more modifications selected from the group consisting of base pair substitutions, base pair insertions, base pair deletions, copy number alterations (CNAs), gene rearrangements, and any combination thereof.

[0043] E13. High TMB is at least 210, at least 215, at least 220, at least 225, at least 230, at least 235, at least 240, at least 245, at least 250, at least 255, at least 260, at least 265, at least 270, at least 275, at least 280, at least 285, at least 290, at least 295, at least 300, at least 305, at least 310, at least 315, at least 320, at least 325, at least 330, at least 335, at least 340, at least 345, at least 350, at least 355, at least 360, at least 365, at least 370, at least 375, at least 380, at least 385, at least 390, at least 400, at least 410, at least 415, at least 420, at least 425, at least 430, at least 435, at least 440, at least 445, at least 450, at least 455, at least 460, at least 470, at least 480, at least 490, at least 500, at least 510, at least 520, at least 530, at least 540, at least 555, at least 560, at least 570, at least 580, at least 590, at least 600, at least 610, at least 620, at least 630, at least 640, at least 650, at least 660, at least 670, at least 680, at least 690 60, at least 365, at least 370, at least 375, at least 380, at least 385, at least 390, at least 395, at least 400, at least 405, at least 410, at least 415, at least 420, at least 425, at least 430, at least 435, at least 440, at least 445, at least 450, at least 455, at least 460, at least 465, at least 470, at least 475, at least 480, at least 485, at least 490, at least 495 or at least 500.

[0044] E14. The method of any of E1-E12, wherein high TMB has a score of at least 215, at least 220, at least 221, at least 222, at least 223, at least 224, at least 225, at least 226, at least 227, at least 228, at least 229, at least 230, at least 231, at least 232, at least 233, at least 234, at least 235, at least 236, at least 237, at least 238, at least 239, at least 240, at least 241, at least 242, at least 243, at least 244, at least 245, at least 246, at least 247, at least 248, at least 249, or at least 250.

[0045] E15. The method of any of E1 to E14, wherein high TMB has a score of at least 243.

[0046] E16. The method of any of E1-E15, further comprising comparing the TMB status of the subject with a control TMB value.

[0047] E17. The method of E16, wherein the subject's TMB status is within the highest quantile of control TMB values.

[0048] E18. The method of E16, wherein the subject's TMB status is within the highest tertile of control TMB values.

[0049] E19. The method according to any one of E1 to E18, wherein the biological sample is a tumor tissue biopsy sample.

[0050] E20. The method according to E19, wherein the tumor tissue is formalin-fixed, paraffin-embedded tumor tissue or fresh-frozen tumor tissue.

[0051] E21. The method of any one of E1 to E18, wherein the biological sample is a liquid biopsy sample.

[0052] E22. The method of any one of E1 to E18, wherein the biological sample comprises one or more of blood, serum, plasma, exoRNA, circulating tumor cells, ctDNA, and cfDNA.

[0053] E23. The method of any one of E1 to E22, wherein TMB status is determined by genome sequencing.

[0054] E24. The method of any of E1 to E22, wherein TMB status is determined by exome sequencing.

[0055] E25. The method of any of E1 to E22, wherein TMB status is determined by genomic profiling.

[0056] E26. The method of E25, wherein the genomic profile comprises at least 300 genes, at least 305 genes, at least 310 genes, at least 315 genes, at least 320 genes, at least 325 genes, at least 330 genes, at least 335 genes, at least 340 genes, at least 345 genes, at least 350 genes, at least 355 genes, at least 360 genes, at least 365 genes, at least 370 genes, at least 375 genes, at least 380 genes, at least 385 genes, at least 390 genes, at least 395 genes, or at least 400 genes.

[0057] E27. The method of E25, wherein the genomic profile comprises at least 325 genes.

[0058] E28. ABL1, BRAF, CHEK1, FANCC, GATA3, JAK2, MITF, PDCD1LG2, RBM10 STAT4, ABL2, BRCA1, CHEK2, FANCD2, GATA4, JAK3, MLH1, PDGFRA, RET, STK1 1, ACVR1B, BRCA2, CIC, FANCE, GATA6, JUN, MPL, PDGFRB, RICTOR, SUFU, AKT1 BRD4, CREBBP, FANCF, GID4(C17orf39), KAT6A(MYST3), MRE11A, PDK1, RNF 43. SYK, AKT2, BRIP1, CRKL, FANCG, GLI1, KDM5A, MSH2, PIK3C2B, ROS1, TAF1 AKT3, BTG1, CRLF2, FANCL, GNA11, KDM5C, MSH6, PIK3CA, RPTOR, TBX3, ALK. BTK, CSF1R, FAS, GNA13, KDM6A, MTOR, PIK3CB, RUNX1, TERC, AMER1(FAM123B). )、C11orf30(EMSY)、CTCF、FAT1、GNAQ、KDR、MUTYH、PIK3CG、RUNX1T1、TERT( APC, CARD11, CTNNA1, FBXW7, GNAS, KEAP1, MYC, PIK3R1, SDHA, T.S ET2, AR, CBFB, CTNNB1, FGF10, GPR124, KEL, MYCL(MYCL1), PIK3R2, SDHB, TG FBR2, ARAF, CBL, CUL3, FGF14, GRIN2A, KIT, MYCN, PLCG2, SDHC, TNFAIP3, AR FRP1, CCND1, CYLD, FGF19, GRM3, KLHL6, MYD88, PMS2, SDHD, TNFRSF14, ARID 1A, CCND2, DAXX, FGF23, GSK3B, KMT2A(MLL), NF1, POLD1, SETD2, TOP1, ARID 1B, CCND3, DDR2, FGF3, H3F3A, KMT2C(MLL3), NF2, POLE, SF3B1, TOP2A, ARID 2. CCNE1, DICER1, FGF4, HGF, KMT2D(MLL2), NFE2L2, PPP2R1A, SLIT2, TP53. ASXL1, CD274, DNMT3A, FGF6, HNF1A, KRAS, NFKBIA, PRDM1, SMAD2, TSC1, ATM.CD79A, DOT1L, FGFR1, HRAS, LMO1, NKX2-1, PREX2, SMAD3, TSC2, ATR, CD79B, EGFR, FGFR2, HSD3B1, LRP1B, NOTCH1, PRKAR1A, SMAD4, TSHR, ATRX, CDC73, EP300, FGFR3, HSP90AA1, LYN, NOTCH2, PRKCI, SMARCA4, U2AF1, AURKA, CDH1, EPHA3, FGFR4, IDH1, LZTR1, NOTCH3, PRKDC, SMARCB1, VEGF A, AURKB, CDK12, EPHA5, FH, IDH2, MAGI2, NPM1, PRSS8, SMO, VHL, AXIN1, CDK4, EPHA7, FLCN, IGF1R, MAP2K1, NRAS, PTCH1, SNCAIP, WISP3, AXL, C DK6, EPHB1, FLT1, IGF2, MAP2K2, NSD1, PTEN, SOCS1, WT1, BAP1, CDK8, ERBB2, FLT3, IKBKE, MAP2K4, NTRK1, PTPN11, SOX10, XPO1, BARD1, CDKN1A, ERBB3, FLT4, IKZF1, MAP3K1, NTRK2, QKI, SOX2, ZBTB2, BCL2, CDKN1B, ERBB4, FOXL2, IL7R, MCL1, NTRK3, RAC1, SOX9, ZNF217, BCL2L1, CDKN2A, E RG, FOXP1, INHBA, MDM2, NUP93, RAD50, SPEN, ZNF703, BCL2L2, CDKN2B, ERRFI1, FRS2, INPP4B, MDM4, PAK3, RAD51, SPOP, BCL6, CDKN2C, ESR1, FU The method according to any one of E25 to E27, wherein the gene comprises one or more genes selected from the group consisting of BP1, IRF2, MED12, PALB2, RAF1, SPTA1, BCOR, CEBPA, EZH2, GABRA6, IRF4, MEF2B, PARK2, RANBP2, SRC, BCORL1, CHD2, FAM46C, GATA1, IRS2, MEN1, PAX5, RARA, STAG2, BLM, CHD4, FANCA, GATA2, JAK1, MET, PBRM1, RB1, STAT3, and any combination thereof.

[0059] E29. The method of any one of E1 to E28, further comprising identifying genomic alterations in one or more of ETV4, TMPRSS2, ETV5, BCR, ETV1, ETV6 and MYB.

[0060] E30. The method of any of E1-E29, wherein the subject has a tumor with a high neoantigen load.

[0061] E31. The method of any of E1-E30, wherein the subject has an expanded T cell repertoire.

[0062] E32. The method according to any one of E1 to E31, wherein the SCLC comprises small cell carcinoma.

[0063] E33. The method according to any one of E1 to E31, wherein the SCLC comprises mixed small cell carcinoma.

[0064] E34. The method according to any one of E1 to E33, wherein the SCLC is recurrent SCLC.

[0065] E35. The method of any of E1-E34, wherein the subject has had at least one, at least two, at least three, at least four, or at least five prior therapies to treat the tumor.

[0066] E36. The method of E35, wherein the prior treatment experience includes chemotherapy.

[0067] E37. The method of E36, wherein the chemotherapy comprises platinum-based therapy.

[0068] E38. The method of E37, wherein the platinum-based therapy comprises a platinum-based anti-neoplastic selected from the group consisting of cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenanthriplatin, picoplatin, satraplatin, and any combination thereof.

[0069] E39. The method of E37 or E38, wherein the platinum-based therapy comprises cisplatin.

[0070] E40. The method of any of E1 to E39, wherein the anti-PD-1 antibody cross-competes with nivolumab for binding to human PD-1.

[0071] E41. The method according to any one of E1 to E40, wherein the anti-PD-1 antibody binds to the same epitope as nivolumab.

[0072] E42. The method of any one of E1 to E41, wherein the anti-PD-1 antibody is a chimeric antibody, a humanized antibody, a human monoclonal antibody, or an antigen-binding portion thereof.

[0073] E43. The method of any one of E1 to E42, wherein the anti-PD-1 antibody comprises a heavy chain constant region of a human IgG1 isotype or a human IgG4 isotype.

[0074] E44. The method according to any one of E1 to E43, wherein the anti-PD-1 antibody is nivolumab.

[0075] E45. The method according to any one of E1 to E43, wherein the anti-PD-1 antibody is pembrolizumab.

[0076] E46. The method of any one of E1 to E45, wherein the anti-PD-1 antibody is administered every 2, 3, or 4 weeks at a dose ranging from 0.1 mg / kg to E10.0 mg / kg body weight.

[0077] E47. The method of any of E1 to E46, wherein the anti-PD-1 antibody is administered at a dose of 5 mg / kg or E10 mg / kg body weight every three weeks.

[0078] E48. The method of any one of E1 to E47, wherein the anti-PD-1 antibody is administered at a dose of 5 mg / kg body weight every three weeks.

[0079] E49. The method of any one of E1 to E46, wherein the anti-PD-1 antibody is administered at a dose of 3 mg / kg body weight every two weeks.

[0080] E50. The method of any of E1-E45, wherein the anti-PD-1 antibody is administered as a fixed dose.

[0081] E51. The method of E50, wherein the anti-PD-1 antibody is administered as a fixed dose of at least about 200 mg, at least about 220 mg, at least about 240 mg, at least about 260 mg, at least about 280 mg, at least about 300 mg, at least about 320 mg, at least about 340 mg, at least about 360 mg, at least about 380 mg, at least about 400 mg, at least about 420 mg, at least about 440 mg, at least about 460 mg, at least about 480 mg, at least about 500 mg, or at least about 550 mg.

[0082] E52. The method of E50 or E51, wherein the anti-PD-1 antibody is administered at a fixed dose about every 1, 2, 3, or 4 weeks.

[0083] E53. The method of any one of E2, E3, E5, and E6 to E52, wherein the anti-CTLA-4 antibody is a chimeric, humanized, or human monoclonal antibody or a portion thereof.

[0084] E54. The method of any of E2, E3, E5, and E6-E53, wherein the anti-CTLA-4 antibody comprises a heavy chain constant region that is of the human IgG1 isotype.

[0085] E55. The method according to any one of E2, E3, E5, and E6 to E55, wherein the anti-CTLA-4 antibody is ipilimumab.

[0086] E56. The method according to any one of E2, E3, E5, and E6 to E55, wherein the anti-CTLA-4 antibody is tremelimumab.

[0087] E57. The method of any of E2, E3, E5, and E6-E56, wherein the anti-CTLA-4 antibody cross-competes with ipilimumab for binding to human CTLA-4.

[0088] E58. The method of any one of E2, E3, E5, and E6 to E57, wherein the anti-CTLA-4 antibody is administered at a dose ranging from at least about 0.1 mg / kg to at least about 10.0 mg / kg body weight approximately every 1, 2, 3, or 4 weeks.

[0089] E59. The method of any of E2, E3, E5, and E6 to E57, wherein the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg or about 3 mg / kg body weight.

[0090] E60. The method of any of E2, E3, E5, and E6-E57, wherein the anti-CTLA-4 antibody is administered at a fixed dose.

[0091] E61. The method of any one of E2, E3, E5, and E6 to E60, wherein the anti-CTLA-4 antibody is administered approximately every two weeks.

[0092] E62. The method of any one of E2, E3, E5, and E6-E60, wherein the anti-CTLA-4 antibody is administered approximately every three weeks.

[0093] E63. The method of any of E2, E3, E5, and E6 to E57, wherein the anti-PD-1 antibody is administered at a dose of about 3 mg / kg body weight approximately every 3 weeks, and the anti-CTLA-4 antibody is administered at a dose of about 1 mg / kg body weight approximately every 3 weeks.

[0094] E64. The method of any of E2, E3, E5, and E6 to E57, wherein the anti-PD-1 antibody is administered at a dose of about 1 mg / kg body weight approximately every 3 weeks, and the anti-CTLA-4 antibody is administered at a dose of about 3 mg / kg body weight approximately every 3 weeks.

[0095] E65. The method of any of E2, E3, E5, and E6 to E57, wherein (i) an anti-PD-1 antibody is administered at a dose of about 1 mg / kg body weight about every 3 weeks and an anti-CTLA-4 antibody is administered at a dose of about 3 mg / kg body weight about every 3 weeks for 4 cycles, followed by (ii) an anti-PD-1 antibody is administered at a dose of about 3 mg / kg body weight about every 2 weeks.

[0096] E66. The method of any of E1-E65, wherein the subject exhibits progression-free survival for at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 1 year, at least about 18 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years after administration.

[0097] E67. The method of any of E1-E66, wherein the subject exhibits an overall survival of at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 1 year, at least about 14 months, at least about 16 months, at least about 18 months, at least about 20 months, at least about 22 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years after administration.

[0098] E68. The method of any of E1-E67, wherein subjects exhibit an objective response rate of at least about 15%, at least about 20%, at least about 25%, at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100%.

[0099] E69. Genomic profile is FOUNDATIONONE® CDX TM The method according to E25, comprising:

[0100] E70. The method of any of E1-E69, wherein the tumor has a TMB of at least about 10 mutations per megabase of the sequenced genome. [Brief explanation of the drawings]

[0101] [Figure 1]Figure 1 is a schematic diagram of a clinical trial protocol for the treatment of SCLC using an anti-PD-1 antibody, e.g., nivolumab, as monotherapy or combination therapy including an anti-PD-1 antibody, e.g., nivolumab, and an anti-CTLA-4 antibody, e.g., ipilimumab. The database was locked on March 30, 2017. Patients (ITT, N=401; based on data from the previous database lock, only patients in the 1 mg / kg nivolumab + 3 mg / kg ipilimumab and 3 mg / kg nivolumab monotherapy cohorts were selected for further progression in SCLC) received nivolumab monotherapy (n=245) or nivolumab + ipilimumab (n=156). DOR = duration of response; EQ-5D = EuroQoL-5 dimension; ORR = objective response rate; OS = overall survival; PD-L1 = programmed cell death ligand 1; PFS = progression-free survival. a Median follow-up 23.3 months; b Median follow-up 28.6 months; c Median follow-up 10.8 months; d Median follow-up 11.2 months.

[0102] [Figure 2] Figure 2 is a schematic diagram illustrating the method and sample flow of exploratory TMB analysis. a For germline sequencing.

[0103] [Figure 3] 3A-3D are graphical representations of progression-free survival (PFS; FIGS. 3A and 3C) and overall survival (OS; FIGS. 3B and 3D) for subjects treated with an anti-PD-1 antibody, e.g., nivolumab, as a monotherapy (FIGS. 3A and 3B) or a combination therapy including an anti-PD-1 antibody, e.g., nivolumab, and an anti-CTLA-4 antibody, e.g., ipilimumab (FIGS. 3C and 3D). PFS and OS for ITT and TMB-evaluable patients are superimposed as shown (FIGS. 3A-3D).

[0104] [Figure 4]Figures 4A-4C are graphical representations of TMB distribution for subjects in the SCLC clinical trial described herein (Figure 4A), consolidated SCLC trial subjects (Figure 4B), and consolidated subjects from prior clinical trials for the treatment of non-small cell lung cancer (Figure 4C).

[0105] [Figure 5] FIG. 5 is a bar graph showing the overall response rate (ORR) for all TMB-evaluable subjects and the same subjects stratified by TMB status (low, intermediate, or high) treated with an anti-PD-1 antibody, e.g., nivolumab, or an anti-PD-1 antibody, e.g., nivolumab, and an anti-CTLA-4 antibody, e.g., ipilimumab.

[0106] [Figure 6] 6A-6B are graphical representations of TMB distribution for subjects treated with an anti-PD-1 antibody, e.g., nivolumab monotherapy (FIG. 6A) or a combination therapy including an anti-PD-1 antibody, e.g., nivolumab, and an anti-CTLA-4 antibody, e.g., ipilimumab (FIG. 6B), where subjects are stratified by best overall response: CR = complete response; PR = partial response; SD = stable disease; PD = progressive disease; NE = not evaluated.

[0107] [Figure 7A] 7A-7B show progression-free survival (PFS) in subjects treated with an anti-PD-1 antibody, e.g., nivolumab, as monotherapy (FIG. 7A) or a combination therapy including an anti-PD-1 antibody, e.g., nivolumab, and an anti-CTLA-4 antibody, e.g., ipilimumab (FIG. 7B), stratified by TMB status (low, intermediate, or high), as indicated. One-year PFS is shown for each sample population. [Figure 7B] Same as above.

[0108] [Figure 8A]Figures 8A-8B show overall survival (OS) in subjects treated with an anti-PD-1 antibody, e.g., nivolumab monotherapy (Figure 8A) or a combination therapy including an anti-PD-1 antibody, e.g., nivolumab, and an anti-CTLA-4 antibody, e.g., ipilimumab (Figure 8B), stratified by TMB status (low, intermediate, or high), as indicated. 1-year OS is shown for each sample population. [Figure 8B] Same as above. DETAILED DESCRIPTION OF THE INVENTION

[0109] Detailed Description of the Invention The present invention relates to a method for treating a patient with small cell lung cancer whose tumor has a high TMB status, comprising administering to the patient an anti-PD-1 antibody monotherapy or a combination therapy comprising an anti-PD-1 antibody and an anti-CTLA-4 antibody. The present invention also relates to a method for identifying a patient with small cell lung cancer who is suitable for treatment with an anti-PD-1 antibody monotherapy or a combination therapy comprising an anti-PD-1 antibody and an anti-CTLA-4 antibody, comprising measuring the TMB status of a biological sample from the patient.

[0110] term So that the present invention may be more readily understood, certain terms are first defined. As used herein, unless expressly stated otherwise, each of the following terms has the meaning indicated below. Additional definitions are set forth throughout the specification.

[0111] "Administration" refers to the physical introduction of a composition containing a therapeutic agent into a subject using any of a variety of methods and delivery systems known to those skilled in the art. Preferred routes of administration for anti-PD-1 antibodies are intravenous, intramuscular, subcutaneous, intraperitoneal, spinal, or other parenteral routes, for example, by injection or infusion. As used herein, the term "parenteral administration" refers to modes of administration other than enteral and topical administration, usually by injection, and includes, but is not limited to, intravenous, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transbronchial, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion, as well as in vivo electroporation. TKIs are generally administered by non-parenteral routes, preferably orally. Other non-parenteral routes include topical, epithelial, or mucosal routes of administration, for example, intranasal, vaginal, rectal, sublingual, or topical. Administration can be carried out, for example, once, multiple times and / or over a longer period of time.

[0112] As used herein, an "adverse event" (AE) is any untoward, generally unintended or undesirable sign (including laboratory abnormalities), symptom, or disease associated with the use of a drug therapy. For example, an adverse event may be associated with activation of the immune system or an increase in immune system cells (e.g., T cells) in response to treatment. A drug therapy may have one or more associated AEs, and the severity level of each AE may be the same or different. References to methods that can "alter an adverse event" refer to a treatment regimen that reduces the incidence and / or severity of one or more AEs associated with the use of a different treatment regimen.

[0113] An "antibody" (Ab) includes, but is not limited to, a glycoprotein immunoglobulin or antigen-binding portion thereof that specifically binds to an antigen and comprises at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each H chain contains a heavy chain variable region (herein referred to as a V H The heavy chain constant region contains three constant domains: C H1 , C H2 and C H3 Each light chain contains a light chain variable region (herein V LThe light chain constant region contains one constant domain, C L Includes V H and V L The regions can be further subdivided into regions of hypervariability, termed complementarity-determining regions (CDRs), interspersed with more conserved regions, termed framework regions (FRs). H and V L contains three CDRs and four FRs, arranged from the amino terminus to the carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The variable regions of the heavy and light chains contain binding domains that interact with antigens. The constant regions of the antibody may mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system.

[0114] Immunoglobulins may be derived from any of the commonly known isotypes, including, but not limited to, IgA, secretory IgA, IgG, and IgM. IgG subclasses are also well known in the art and include, but are not limited to, human IgG1, IgG2, IgG3, and IgG4. "Isotype" refers to the antibody class or subclass (e.g., IgM or IgG1) encoded by heavy chain constant region genes. The term "antibody" includes, by way of example, both naturally occurring and non-naturally occurring antibodies; monoclonal and polyclonal antibodies; chimeric and humanized antibodies; human or non-human antibodies; fully synthetic antibodies; and single-chain antibodies. Non-human antibodies may be recombinantly humanized to reduce immunogenicity in humans. Unless expressly stated and consistent with the context, the term "antibody" also includes antigen-binding fragments or portions of any of the above immunoglobulins, including monovalent and bivalent fragments or portions and single-chain antibodies.

[0115] An "isolated antibody" refers to an antibody that is substantially free of other antibodies having different antigenic specificities (e.g., an isolated antibody that specifically binds PD-1 is substantially free of antibodies that specifically bind to antigens other than PD-1). However, an isolated antibody that specifically binds PD-1 may have cross-reactivity with other antigens, such as PD-1 molecules from different species. Moreover, an isolated antibody may be substantially free of other cellular material and / or chemicals.

[0116] The term "monoclonal antibody" (mAb) refers to a non-naturally occurring preparation of antibody molecules of single molecular composition, i.e., antibody molecules which are substantially identical in primary sequence and which display a single binding specificity and affinity for a particular epitope. A monoclonal antibody is an example of an isolated antibody. Monoclonal antibodies may be produced by hybridoma, recombinant, transgenic, or other techniques known to those skilled in the art.

[0117] A "human antibody" (HuMAb) refers to an antibody having variable regions in which both the framework and CDR regions are derived from human germline immunoglobulin sequences. Furthermore, if the antibody contains a constant region, the constant region also is derived from human germline immunoglobulin sequences. The human antibodies of the invention may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo). However, as used herein, the term "human antibody" is not intended to include antibodies in which CDR sequences derived from the germline of other mammalian species, such as a mouse, have been grafted onto human framework sequences. The terms "human antibody" and "fully human antibody" are used interchangeably.

[0118] A "humanized antibody" refers to an antibody in which some, most, or all of the amino acids outside the CDRs of a non-human antibody have been replaced with corresponding amino acids from a human immunoglobulin. In some embodiments of a humanized form of an antibody, some, most, or all of the amino acids outside the CDRs have been replaced with amino acids from a human immunoglobulin, while some, most, or all of the amino acids within one or more CDRs remain unchanged. Small additions, deletions, insertions, substitutions, or modifications of amino acids are permissible as long as they do not abrogate the ability of the antibody to bind to a specific antigen. A "humanized antibody" retains antigen specificity similar to that of the original antibody.

[0119] "Chimeric antibody" refers to an antibody in which the variable region is derived from one species and the constant region is derived from another species, such as an antibody in which the variable region is derived from a mouse antibody and the constant region is derived from a human antibody.

[0120] An "anti-antigen antibody" refers to an antibody that specifically binds to the antigen. For example, an anti-PD-1 antibody specifically binds to PD-1.

[0121] An "antigen-binding portion" (also called an "antigen-binding fragment") of an antibody refers to one or more fragments of an antibody that retain the ability to specifically bind to the antigen bound by the whole antibody.

[0122] "Cancer" refers to a wide group of diverse diseases characterized by the uncontrolled growth of abnormal cells in the body. Uncontrolled cell division and proliferation leads to the formation of malignant tumors that can invade neighboring tissues and even metastasize to distant parts of the body through the lymphatic system or bloodstream.

[0123] The term "immunotherapy" refers to the treatment of a subject having or at risk of developing or recurring a disease by a method that induces, enhances, suppresses, or otherwise modifies the immune response. "Treatment" or "treatment" of a subject refers to any type of intervention or procedure or administration of an active agent performed on a subject with the intent to reverse, alleviate, ameliorate, arrest, slow down, or prevent the onset, progression, development, severity, or recurrence of a symptom, complication, or condition, or biochemical marker associated with a disease.

[0124] "Programmed cell death-1" (PD-1) refers to an immunoinhibitory receptor belonging to the CD28 family. PD-1 is primarily expressed on pre-activated T cells in vivo and binds to two ligands, PD-L1 and PD-L2. As used herein, the term "PD-1" includes human PD-1 (hPD-1), variants, isoforms, and species homologs of hPD-1, as well as analogs that share at least one epitope with hPD-1. The complete hPD-1 sequence can be found under GenBank Accession No. U64863.

[0125] "Programmed death-ligand-1" (PD-L1) is one of two cell surface glycoprotein ligands for PD-1 (the other being PD-L2) that downregulates T cell activation and cytokine secretion upon binding to PD-1. As used herein, the term "PD-L1" includes human PD-L1 (hPD-L1), variants, isoforms, and species homologs of hPD-L1, as well as analogs that share at least one epitope with hPD-L1. The complete hPD-L1 sequence can be found under GenBank Accession No. Q9NZQ7.

[0126] A "subject" includes any human or non-human animal. The term "non-human animal" includes, but is not limited to, vertebrates such as non-human primates, sheep, dogs, and rodents, such as mice, rats, and guinea pigs. In a preferred embodiment, the subject is a human. The terms "subject" and "patient" are used interchangeably herein.

[0127] The use of the term "fixed dose" in connection with the methods and dosages of the invention refers to a dose administered to a patient without regard to the patient's weight or body surface area (BSA). A fixed dose is therefore provided as an absolute amount of agent (e.g., an anti-PD-1 antibody) rather than a mg / kg dose. For example, a 60 kg human and a 100 kg human may receive the same dose of antibody (e.g., 240 mg of an anti-PD-1 antibody). In one embodiment, an anti-PD-1 antibody is administered in the methods described herein at a fixed dose of about 240 mg every two weeks. In one embodiment, an anti-PD-1 antibody is administered in the methods described herein at a fixed dose of about 480 mg every four weeks. In another embodiment, an anti-PD-1 antibody is administered in the methods described herein at a fixed dose of about 200 mg every three weeks.

[0128] The use of the term "fixed dose" in connection with the methods of the present invention means that two or more different antibodies (e.g., an anti-PD-1 antibody and an anti-CTLA-4 antibody) are present in a single composition, particularly in a (fixed) ratio to each other. In certain embodiments, the fixed dose is based on the weight (e.g., mg) of the antibody. In certain embodiments, the fixed dose is based on the concentration (e.g., mg / ml) of the antibody. In certain embodiments, the ratio is at least about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, about 1:30, about 1:40, about 1:50, about 1:60, about 1:70, about 1:80, about 1:90, about 1:100, about 1:120, about 1:140, about 1:160, about 1:180, about 1:200, about 200:1, about 200:1, about 200:1 , about 160:1, about 140:1, about 120:1, about 100:1, about 90:1, about 80:1, about 70:1, about 60:1, about 50:1, about 40:1, about 30:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, or about 2:1 mg of first antibody (e.g., anti-PD-1 antibody) to mg of second antibody (e.g., anti-CTLA-4 antibody). For example, a 3:1 ratio of anti-PD-1 antibody and anti-CTLA-4 antibody means that a vial can contain about 240 mg of anti-PD-1 antibody and 80 mg of anti-CTLA-4 antibody, or about 3 mg / ml of anti-PD-1 antibody and 1 mg / ml of anti-CTLA-4 antibody.

[0129] As used herein, the term "weight-based dose" means that the dose administered to a patient is calculated based on the patient's weight. For example, if a patient weighing 60 kg requires 3 mg / kg of anti-PD-1 antibody, an appropriate amount of anti-PD-1 antibody (i.e., 180 mg) can be calculated and used for administration.

[0130] A "therapeutically effective amount" or "therapeutically effective dose" of a drug or therapeutic agent is any amount of drug that, when used alone or in combination with other therapeutic agents, promotes disease regression that protects a subject from developing the disease, as evidenced by a reduction in the severity of disease symptoms, an increase in the frequency and duration of disease-free symptom intervals, or prevention of functional impairment or disability due to disease morbidity. The ability of a therapeutic agent to promote disease regression can be assessed by a variety of methods known to the skilled practitioner, such as by assaying the activity of the agent in human subjects in clinical trials, in animal model systems predictive of efficacy in humans, or in in vitro assays.

[0131] By way of example, an "anti-cancer agent" promotes cancer regression in a subject. In a preferred embodiment, a therapeutically effective amount of a drug promotes cancer regression to the point of eliminating the cancer. "Promoting cancer regression" means that administration of an effective amount of a drug, alone or in combination with an anti-neoplastic agent, results in a reduction in tumor growth or size, tumor necrosis, a reduction in the severity of at least one disease symptom, an increase in the frequency and duration of disease-free symptom intervals, or prevention of functional impairment or disability due to disease. Furthermore, the terms "effective" and "efficacy" in relation to treatment include both pharmacological effectiveness and physiological safety. Pharmacological effectiveness refers to the ability of a drug to promote cancer regression in a patient. Physiological safety refers to the level of toxicity or other adverse physiological effects (adverse effects) at the cellular, organ, and / or organismal level resulting from drug administration.

[0132] For example, in the context of tumor treatment, a therapeutically effective amount of an anticancer agent preferably inhibits cell proliferation or tumor growth by at least about 20%, more preferably at least about 40%, even more preferably at least about 60%, and even more preferably at least about 80% compared to untreated subjects. In other preferred embodiments of the present invention, tumor regression can be observed and continued for a period of at least about 20 days, more preferably at least about 40 days, or even more preferably at least about 60 days. Regardless of these final assessments of therapeutic efficacy, the evaluation of immunotherapy drugs should also take into account immune-related response patterns.

[0133] "Immune response," as understood in the art, generally refers to a biological response in a vertebrate to foreign agents or abnormalities, e.g., cancer cells, which protects the organism from these agents and the diseases they cause. An immune response is an action mediated by one or more cells of the immune system (e.g., T lymphocytes, B lymphocytes, natural killer (NK) cells, macrophages, eosinophils, mast cells, dendritic cells, or neutrophils) and soluble macromolecules (including antibodies, cytokines, and complement) produced by any of these cells or the liver, which results in the selective targeting, binding, damaging, destruction, and / or elimination of invading pathogens, pathogen-infected cells or tissues, cancer or other abnormal cells, or, in the case of autoimmunity or pathological inflammation, normal human cells or tissues of the vertebrate body. An immune response can be mediated by T cells, e.g., effector T cells, Th cells, CD4 + cells, CD8 + This includes, for example, the activation or inhibition of T cells or Treg cells or the activation or inhibition of any other cells of the immune system, for example, NK cells.

[0134] "Immune-related response pattern" refers to a clinical response pattern often observed in cancer patients treated with immunotherapeutic agents that produce antitumor effects by eliciting cancer-specific immune responses or modifying natural immune processes. This response pattern is characterized by a favorable therapeutic effect after an increase in tumor burden or the appearance of new lesions, which would be classified as disease progression in the evaluation of traditional chemotherapeutic agents and is synonymous with drug failure. Therefore, proper evaluation of immunotherapeutic agents may require long-term monitoring of the effects of these agents on the target disease.

[0135] "Immunomodulatory agent" or "immunoregulatory agent" refers to an agent, e.g., an agent that targets a component of a signal transduction pathway that may be involved in the regulation, regulation, or modification of an immune response. "Modulation," "regulation," or "modulation" of an immune response refers to any alteration of the cells of the immune system or the activity of such cells (e.g., effector T cells, such as Th1 cells). Such modulation includes stimulation or suppression of the immune system, which may be manifested by an increase or decrease in the number of various cell types, an increase or decrease in the activity of these cells, or any other change that may occur within the immune system. Both inhibitory and stimulatory immunomodulatory agents have been identified, some of which may have enhanced function in the tumor microenvironment. In certain embodiments, an immunomodulatory agent targets a molecule on the surface of T cells. An "immunoregulatory target" or "immunoregulatory target" is a molecule, e.g., a cell surface molecule, that is the target of binding of a substance, agent, moiety, compound, or molecule, and whose activity is altered upon binding. Immunoregulatory targets include, for example, receptors on the cell surface ("immunoregulatory receptors") and receptor ligands ("immunoregulatory ligands").

[0136] "Immunotherapy" refers to the treatment of a subject having a disease or at risk of developing or recurring by methods that involve eliciting, enhancing, suppressing, or otherwise modifying the immune system or immune response. In some embodiments, immunotherapy involves the administration of an antibody to a subject. In other embodiments, immunotherapy refers to the administration of a small molecule to a subject. In other embodiments, immunotherapy involves the administration of a cytokine or an analog, variant, or fragment thereof.

[0137] "Immunostimulatory therapy" or "immunostimulatory therapy" refers to a therapy that results in an increase (induction or enhancement) of the immune response in a subject, for example, to treat cancer.

[0138] "Endogenous enhancement" means increasing the efficacy or potency of an existing immune response in a subject. This increased efficacy and potency can be achieved, for example, by overcoming mechanisms that suppress the endogenous host immune response or by stimulating mechanisms that enhance the endogenous host immune response.

[0139] A therapeutically effective amount of a drug includes a "prophylactically effective amount," which is any amount of drug that, when administered alone or in combination with an anti-neoplastic agent to a subject at risk of developing cancer (e.g., a subject with a pre-malignant condition) or having cancer recurrence, prevents the onset or recurrence of cancer. In preferred embodiments, a prophylactically effective amount completely prevents the onset or recurrence of cancer. "Preventing" the onset or recurrence of cancer means reducing the likelihood of cancer onset or recurrence or completely preventing the onset or recurrence of cancer.

[0140] As used herein, the term "tumor mutation burden" (TMB) refers to the number of somatic mutations in a tumor's genome and / or the number of somatic mutations per region of the tumor's genome. Germline (inherited) variants are excluded when determining TMB because the immune system is more likely to recognize them as self. Tumor mutation burden (TMB) is also used interchangeably with "tumor mutation load," "tumor mutational burden," or "tumor mutational load."

[0141] TMB is a genetic analysis of the tumor genome and can therefore be measured by applying sequencing methods well known to those skilled in the art. Tumor DNA can be compared with DNA from patient-matched normal tissue to exclude germline mutations or polymorphisms.

[0142] In some embodiments, TMB is determined by sequencing tumor DNA using high-throughput sequencing technology, such as next-generation sequencing (NGS) or NGS-based methods. In some embodiments, the NGS-based method is whole genome sequencing (WGS), whole exome sequencing (WES), or FOUNDATIONONE CDX. TMand comprehensive genomic profiling (CGP) of cancer gene panels, such as the MSK-IMPACT clinical trial. In some embodiments, TMB as used herein refers to the number of somatic mutations per megabase pair (Mb) of sequenced DNA. In some embodiments, TMB is measured by using the total number of non-synonymous mutations, such as missense mutations (i.e., altering specific amino acids in proteins) and / or nonsense mutations (leading to premature termination and therefore shortening of protein sequences), identified by standardizing with matched tumor germline samples to eliminate any inherited germline genetic alterations. In other embodiments, TMB is measured by using the total number of missense mutations in tumors. To measure TMB, a sufficient amount of sample is required. In some embodiments, tissue samples (e.g., at least 10 slides) are used for evaluation. In some embodiments, TMB is expressed as NsM per megabase pair (NsM / Mb). One megabase pair represents 1 million base pairs.

[0143] TMB status can be a numerical or relative value, eg, high, medium, or low; within the highest quantile or within the highest tertile of a control set.

[0144] As used herein, the term "high TMB" refers to a number of somatic mutations in the genome of a tumor that is above normal or average. In some embodiments, the TMB is at least 210, at least 215, at least 220, at least 225, at least 230, at least 235, at least 240, at least 245, at least 250, at least 255, at least 260, at least 265, at least 270, at least 275, at least 280, at least 285, at least 290, at least 295, at least 300, at least 305, at least 310, at least 315, at least 320, at least 325, at least 330, at least 335, at least 340, at least 345, at least 350, at least 355, at least 360, at least 365, at least 370, at least 375, at least 380, at least 385, at least 390, at least 395, at least 400, at least 405, at least 410, at least 415, at least 420, at least 425, at least 430, at least 435, at least 440, at least 445 In certain embodiments, a high TMB has a score of at least 45, 450, at least 455, at least 460, at least 465, at least 470, at least 475, at least 480, at least 485, at least 490, at least 495, or at least 500; in other embodiments, a high TMB has a score of at least 221, at least 222, at least 223, at least 224, at least 225, at least 226, at least 227, at least 228, at least 229, at least 230, at least 231, at least 232, at least 233, at least 234, at least 235, at least 236, at least 237, at least 238, at least 239, at least 240, at least 241, at least 242, at least 243, at least 244, at least 245, at least 246, at least 247, at least 248, at least 249, or at least 250; and in certain embodiments, a high TMB has a score of at least 243. In other embodiments, "high TMB" refers to TMB within the highest quantile of control TMB values.For example, all subjects with assessable TMB data are classified by TMB quantile distribution, i.e., subjects are ranked from the highest number of genetic alterations to the lowest number, and divided into a specified number of groups.In some embodiments, all subjects with assessable TMB data are ranked and divided into three groups, and " high TMB " is within the highest tertile of control TMB values.In certain embodiments, the tertile boundaries are 0<100 genetic alterations; 100~243 genetic alterations; and >243 genetic alterations.It should be understood that once ranked, subjects with assessable TMB data can be divided into any number of groups, for example, quartiles, quintiles, etc. In certain embodiments, "high TMB" refers to a TMB of at least about 20 mutations / tumor, at least about 25 mutations / tumor, at least about 30 mutations / tumor, at least about 35 mutations / tumor, at least about 40 mutations / tumor, at least about 45 mutations / tumor, at least about 50 mutations / tumor, at least about 55 mutations / tumor, at least about 60 mutations / tumor, at least about 65 mutations / tumor, at least about 70 mutations / tumor, at least about 75 mutations / tumor, at least about 80 mutations / tumor, at least about 85 mutations / tumor, at least about 90 mutations / tumor, at least about 95 mutations / tumor, or at least about 100 mutations / tumor. In certain embodiments, "high TMB" refers to a TMB of at least about 105 mutations / tumor, at least about 110 mutations / tumor, at least about 115 mutations / tumor, at least about 120 mutations / tumor, at least about 125 mutations / tumor, at least about 130 mutations / tumor, at least about 135 mutations / tumor, at least about 140 mutations / tumor, at least about 145 mutations / tumor, at least about 150 mutations / tumor, at least about 175 mutations / tumor, or at least about 200 mutations / tumor. In certain embodiments, a tumor with high TMB has at least about 100 mutations / tumor.

[0145] "High TMB" also refers to, for example, mutation assays, e.g., FOUNDATIONONE CDX TM It may also be referred to as the number of mutations per megabase pair of the sequenced genome as measured by the assay. In some embodiments, high TMB is measured by FOUNDATIONONE CDX TM"High TMB" refers to at least about 9, at least about 10, at least about 11, at least 12, at least about 13, at least about 14, at least about 15, at least about 16, at least about 17, at least about 18, at least about 19, or at least about 20 mutations per megabase pair of the genome as measured by an assay. In certain embodiments, "high TMB" refers to at least about 9, at least about 10, at least about 11, at least about 12, at least about 13, at least about 14, at least about 15, at least about 16, at least about 17, at least about 18, at least about 19, or at least about 20 mutations per megabase pair of the genome as measured by an assay. TM At least 10 mutations per megabase pair of genome sequenced in the assay.

[0146] As used herein, the term "intermediate TMB" refers to a tumor genome with a normal or average or approximately normal or average number of somatic mutations, and the term "low TMB" refers to a tumor genome with a normal or below average number of somatic mutations. In certain embodiments, a "high TMB" has a score of at least 243, a "intermediate TMB" has a score of 100 to 242, and a "low TMB" has a score of less than 100 (or 0 to 100). "Intermediate or low TMB" refers to a tumor genome with a normal or average number of somatic mutations, and a tumor genome with a normal or below average number of somatic mutations. In certain embodiments, a "high TMB" has a score of at least 243, a "intermediate TMB" has a score of 100 to 242, and a "low TMB" has a score of less than 100 (or 0 to 100). "Intermediate or low TMB" refers to a tumor genome with a normal or below average number of somatic mutations, and a tumor genome with a normal or below average number of somatic mutations. TM It has fewer than 9 mutations per megabase pair of the genome sequenced, as determined by the assay.

[0147] As used herein, the term "control TMB value" may be the TMB value shown in Table 9.

[0148] In certain embodiments, TMB status may also be correlated with smoking status. In particular, current or former smokers often have more genetic alterations, e.g., missense mutations, than never-smokers.

[0149] Tumors with high TMB may also have a high neoantigen load. As used herein, the term "neoantigen" refers to a newly formed antigen not previously recognized by the immune system. A neoantigen can be a protein or peptide recognized as foreign (or non-self) by the immune system. Transcription of genes in the tumor genome harboring somatic mutations results in mutant mRNA, which, when translated, generates a mutant protein that is then processed and transported to the ER lumen, binds to the MHC class I complex, and promotes T cell recognition of the neoantigen. Neoantigen recognition can promote T cell activation, clonal expansion, and differentiation into effector and memory T cells. Neoantigen load can correlate with TMB. In some embodiments, TMB is assessed as a surrogate for measuring tumor neoantigen load. The TMB status of a tumor can be used, alone or in combination with other factors, as a factor in determining whether a patient will benefit from a particular anticancer agent or a particular type of treatment or therapy, such as a cancer immunotherapeutic agent, for example, an anti-PD-1 antibody or an antigen-binding portion thereof or an anti-PD-L1 antibody or an antigen-binding portion thereof. In certain embodiments, high TMB status (or high TMB) indicates a high likelihood of benefiting from cancer immunization and can therefore be used to identify patients more likely to benefit from treatment with an anti-PD-1 antibody, or antigen-binding portion thereof. Similarly, tumors with high tumor neoantigen load and high TMB, e.g., from SCLC, are more likely to be immunogenic than tumors with low neoantigen load and low TMB. Furthermore, high neoantigen / high TMB tumors, e.g., from SCLC, are more likely to be recognized as non-self by the immune system and therefore more likely to elicit an immune-mediated anti-tumor response. In certain embodiments, high TMB status and high neoantigen load indicate a high likelihood of benefiting from cancer immunization, e.g., using immunotherapy. As used herein, the term "benefiting from treatment" refers to an improvement in one or more of overall survival, progression-free survival, partial response, complete response, and overall response rate, and also includes a reduction in tumor growth or size, a reduction in the severity of disease symptoms, an increase in the frequency and duration of disease-free symptom intervals, or prevention of functional impairment or disability due to disease.

[0150] Other factors, such as environmental factors, may also be associated with TMB status. For example, smoking status in patients with NSCLC correlates with TMB distribution, with current and former smokers having higher median TMB compared with never-smokers. See Peters et al., AACR, April 1-5, 2017, Washington, DC. Driver mutations in NSCLC tumors were also associated with younger age, female gender, and never-smoker status. See Singal et al., ASCO, June 1-5, 2017; Chicago, IL. A trend was observed correlating lower TMB with the presence of driver mutations such as EGFR, ALK, or KRAS (P=0.06). Davis et al., AACR, April 1-5, 2017, Washington, DC.

[0151] As used herein, the term "somatic mutation" refers to an acquired alteration of DNA that occurs after conception. Somatic mutations can occur in any cell of the body other than germ cells (sperm and eggs) and therefore are not inherited by offspring. These alterations can, but do not always, cause cancer or other diseases. The term "germline mutation" refers to a genetic change in the body's germ cells (egg or sperm) that becomes incorporated into the DNA of all cells in the offspring's body. Germline mutations are passed from parents to offspring. They are also referred to as "inherited mutations." In analyzing TMB, germline mutations are considered the "baseline" and are subtracted from the number of mutations found in tumor biopsies to determine TMB in tumors, e.g., tumors derived from SCLC. Because germline mutations are found in all cells of the body, their presence can be determined by sampling, such as blood or saliva, which is less invasive than tumor biopsy. Germline mutations may increase the risk of developing certain cancers and may play a role in response to chemotherapy.

[0152] The term "measuring" or "measurement," when associated with TMB status, refers to determining a measurable amount of somatic mutations in a subject's biological sample. It is recognized that measurement can be performed by sequencing nucleic acids in the sample, such as cDNA, mRNA, exoRNA, ctDNA, and cfDNA. Measurement can be performed on the subject's sample and / or one or more control samples, and can be, for example, newly detected or correspond to a previous determination. Measurement can be performed by, for example, PCR, qPCR, Sanger sequencing, genomic profiling (including comprehensive gene panels), exome sequencing, genome sequencing, and / or any other method disclosed herein, as known to those skilled in the art. In some embodiments, measurement identifies genomic alterations in the sequenced nucleic acid. Genomic (or gene) profiling methods can include a predetermined set of genes, e.g., a panel of 150-500 genes, and in some cases, genomic alterations assessed in the gene panel are correlated with the total somatic mutations assessed.

[0153] As used herein, the term " genomic alteration " refers to the change (or mutation) in the nucleotide sequence of tumor genome, which does not exist in germline nucleotide sequence, and in some embodiments, it is non-synonymous mutation, including but not limited to base pair substitution, base pair insertion, base pair deletion, copy number change (CNA), gene rearrangement and any combination thereof.In certain embodiments, the genomic alteration measured in biological sample is missense mutation.

[0154] As used herein, the term "whole genome sequencing" or "WGS" refers to a method of sequencing the entire genome. As used herein, the term "whole exome sequencing" or "WES" refers to a method of sequencing all of the protein-coding regions (exons) of the genome.

[0155] As used herein, a "cancer gene panel," "hereditary cancer panel," "comprehensive cancer panel," or "multigene cancer panel" refers to a method of sequencing a subset of target cancer genes. In some embodiments, the CGP includes sequencing at least about 15, at least about 20, at least about 25, at least about 30, at least about 35, at least about 40, at least about 45, or at least about 50 target cancer genes.

[0156] The terms "genomic profiling assay," "global genomic profiling," or "CGP" refer to an assay that analyzes a panel of genes and selects introns for in vitro diagnosis. CGP is a combination of NGS and targeted bioinformatics analysis to screen for mutations in known clinically relevant cancer genes. This method can be used to capture mutations missed by "hotspot" tests (e.g., BRCA1 / BRCA2 mutations or microsatellite markers). In some embodiments, the genes in the panel are cancer-associated genes. In other embodiments, the genomic profiling assay is performed using FOUNDATION ONE (登録商標) It is an assay.

[0157] The term "harmonisation" refers to a study conducted to determine the equivalence between two or more measurements and / or diagnostic tests. Harmonisation studies provide a systematic approach to addressing questions about how multiple diagnostic tests compare to each other and their interchangeability when used to determine a patient's tumor biomarker status. Generally, at least one well-characterised measurement and / or diagnostic test is used as the standard for comparison. Equivalence assessments are often used in harmonisation studies.

[0158] As used herein, the term "concordance" refers to the degree of agreement between two measurements and / or diagnostic tests. Equivalence can be established using both qualitative and quantitative methods. Quantitative methods for assessing equivalence vary based on the type of measurement. A particular measurement may be expressed as 1) a categorical / dichotomous variable or 2) a continuous variable. "Categorical / dichotomous variables" (e.g., above or below the TMB cutoff) may use a concordance rate, such as overall concordance rate (OPA), positive concordance rate (PPA), or negative concordance rate (NPA), to assess equivalence. "Continuous variables" (e.g., TMB by WES) use Spearman's rank correlation or Pearson's correlation coefficient (r), which spans a spectrum of values, with values ​​of -1≦r≦+1 to assess equivalence (note: r=+1 or -1 means that the variables are perfectly correlated). The term "analytical equivalence" refers to the degree of agreement in the performance of two assays or diagnostic tests (e.g., identification of biomarkers, genomic alteration types and genomic signatures, and test reproducibility assessment) to support clinical use. The term "clinical equivalence" refers to the degree of agreement in how two assays or diagnostic tests correlate with clinical outcomes.

[0159] The term "microsatellite instability" or "MSI" refers to a change in the DNA of certain cells (e.g., tumor cells) in which the number of microsatellite repeats (short, repetitive sequences of DNA) differs from the number of repeats present in the inherited DNA. MSI can be high microsatellite instability (MSI-H) or low microsatellite instability (MSI-L). Microsatellites are short, tandem DNA repeats of 1 to 6 base pairs. They are prone to DNA replication errors, which are repaired by mismatch repair (MMR). Therefore, microsatellites are good indicators of genomic instability, particularly defective mismatch repair (dMMR). MSI is usually diagnosed by screening five microsatellite markers (BAT-25, BAT-26, NR21, NR24, and NR27). MSI-H indicates the presence of unstable markers in at least two of the five microsatellite markers analyzed (or ≥30% of the markers when using a large panel). MSI-L means instability of one MSI marker (or 10%-30% of markers in a large panel). MSS means the absence of unstable microsatellite markers.

[0160] As used herein, the term "biological sample" refers to a biological material isolated from a subject. Biological samples can include any biological material suitable for, for example, determining TMB by sequencing nucleic acid in tumor (or circulating tumor cells) and identifying genomic alterations in the sequenced nucleic acid. Biological samples can be any suitable biological tissue or body fluid, such as tumor tissue, blood, plasma and serum. In some embodiments, the sample is a tumor tissue biopsy, such as formalin-fixed, paraffin-embedded (FFPE) tumor tissue or fresh-frozen tumor tissue. In other embodiments, the biological sample is a liquid biopsy, which in some embodiments includes one or more of blood, serum, plasma, circulating tumor cells, exoRNA, ctDNA and cfDNA.

[0161] As used herein, the terms "about every week," "about every 2 weeks," or any other similar administration interval refer to approximate values. "About every week" can include every 7 days ± 1 day, i.e., every 6 to 8 days. "About every 2 weeks" can include every 14 days ± 3 days, i.e., every 11 to 17 days. For example, similar approximations apply to about every 3 weeks, about every 4 weeks, about every 5 weeks, about every 6 weeks, and about every 12 weeks. In certain embodiments, an administration interval of about every 6 weeks or about every 12 weeks means that the first dose may be administered on any day of the week in the first week, and the subsequent dose may be administered on any day of the week in the sixth or twelfth week, respectively. In other embodiments, an administration interval of about every 6 weeks or about every 12 weeks means that the first dose is administered on a specific day of the week (e.g., Monday), and then the subsequent dose is administered on the same day of the week (i.e., Monday) in the sixth or twelfth week, respectively.

[0162] The use of the alternative (e.g., "or") should be understood to mean either one, both, or any combination thereof of the alternatives. As used herein, the singular "a," "an," or "the" should be understood to refer to "one or more" of any described or listed elements.

[0163] The terms "about" or "essentially comprising" refer to a value or composition that is within an acceptable error range for a particular value or composition as determined by one of ordinary skill in the art, which depends in part on how the value or composition is measured or determined, i.e., the limitations of the measurement system. For example, "about" or "essentially comprising" can mean 1 or more standard deviations, as is customary in the art. Alternatively, "about" or "essentially comprising" can mean a range of up to 10%. Furthermore, particularly in biological systems or processes, the term can mean up to an order of magnitude or up to 5 times the value. When a particular value or composition is provided in the specification and claims, unless otherwise specified, the meaning of "about" or "essentially comprising" is presumed to be within an acceptable error range for that particular value or composition.

[0164] As used herein, any concentration range, percentage range, ratio range, or integer range is understood to include every integer value within the stated range and, where appropriate, fractional values ​​thereof (e.g., 1 / 10 and 1 / 100 of an integer), unless otherwise specified.

[0165] A list of abbreviations is provided in Table 1. [Table 1-1] [Table 1-2]

[0166] Various aspects of the invention are now described in further detail.

[0167] Methods of the Invention One embodiment of the present invention relates to a method for identifying a subject having a tumor derived from SCLC and suitable for treatment with an anti-PD-1 antibody, or an antigen-binding portion thereof ("anti-PD-1 antibody") or an anti-PD-L1 antibody, or an antigen-binding portion thereof ("anti-PD-L1 antibody"), comprising measuring the tumor mutational burden (TMB) status of a biological sample from the subject. Another embodiment of the present invention relates to a method for identifying a subject having a tumor derived from SCLC and suitable for treatment with an anti-PD-1 antibody and an anti-CTLA-4 antibody, comprising measuring the tumor mutational burden (TMB) status of a biological sample from the subject.

[0168] As tumors grow, they accumulate somatic mutations that are not present in germline DNA. Tumor mutation burden (TMB) refers to the number of somatic mutations in a tumor's genome and / or the number of somatic mutations per region of the tumor genome (after accounting for germline variant DNA). Somatic mutations, and therefore the acquisition of high TMB, can be influenced by different mechanisms, such as exposure to exogenous mutagens (e.g., smoking or UV light exposure) and DNA mismatch repair mutations (e.g., MSI in colorectal and esophageal cancer). In solid tumors, approximately 95% of mutations are single-base substitutions (Vogelstein et al., Science (2013) 339:1546-1558). A "nonsynonymous mutation" here refers to a nucleotide mutation that changes the amino acid sequence of a protein. Both missense and nonsense mutations can be nonsynonymous mutations. A "missense mutation" here refers to a nonsynonymous point mutation in which a single nucleotide change results in a codon encoding a different amino acid. A "nonsense mutation" herein refers to a nonsynonymous point mutation that changes a codon to a premature stop codon, leading to a truncation of the resulting protein.

[0169] In some embodiments, somatic mutations can be expressed at the RNA and / or protein level, resulting in neoantigens (also referred to as neoepitopes).Neoantigens can affect immune-mediated anti-tumor response.For example, neoantigen recognition can promote T cell activation, clonal proliferation and differentiation into effector and memory T cells.

[0170] As tumors progress, early clonal mutations (or "common mutations") may be present in most or all tumor cells, whereas later mutations (or "branch mutations") may arise in only a subset or region of tumor cells (Yap et al., Sci Tranl Med (2012) 4:1-5; Jamai-Hanjani et al., (2015) Clin Cancer Res 21:1258-1266). As a result, neoantigens derived from clonal "common" mutations are more widespread in the tumor genome than "branch" mutations, and thus, large numbers of T cells may become reactive against clonal neoantigens (McGranahan et al., (2016) 351:1463-1469). In general, tumors with high TMB also have a high neoantigen load, which can lead to high tumor immunogenicity and increased T cell reactivity and antitumor responses. That is, cancers with high TMB may respond well to treatment with immunotherapeutic agents, such as anti-PD-1 or anti-PD-L1 antibodies.

[0171] Advances in sequencing technology allow for the assessment of tumor genomic mutational landscapes. Any sequencing method known in the art can be used to sequence nucleic acids from tumor genomes (e.g., obtained from biological samples of tumor-bearing subjects). In some embodiments, PCR or qPCR, Sanger sequencing, or next-generation sequencing ("NGS") methods (e.g., genomic profiling, exome sequencing, or genome sequencing) can be used to measure TMB. In some embodiments, TMB status is measured using genomic profiling. Genomic profiling involves the analysis of nucleic acids from tumor samples, including coding and non-coding regions, and can be performed using methods with integrated and optimized nucleic acid selection, read alignment, and mutation calling. In some embodiments, gene profiling provides next-generation sequencing (NGS)-based analysis of tumors, which can be optimized on a cancer-by-cancer, gene-by-gene, and / or site-by-site basis. Genomic profiling can integrate the use of multiple, individually tailored alignment methods or algorithms to optimize the performance of sequencing methods, especially those that utilize the massively parallel sequencing of multiple diverse genetic events in multiple diverse genes.Genomic profiling provides for the comprehensive analysis of target cancer genome with clinical-grade quality, and genetic analysis results can be applied to relevant scientific and medical knowledge to improve the quality and efficiency of cancer treatment.

[0172] Genomic profiling involves a panel of genes with a predetermined set, including as few as 5 genes or as many as 1,000 genes, from about 25 to about 750 genes, from about 100 to about 800 genes, from about 150 to about 500 genes, from about 200 to about 400 genes, or from about 250 to about 350 genes. In certain embodiments, the genomic profile includes at least 300 genes, at least 305 genes, at least 310 genes, at least 315 genes, at least 320 genes, at least 325 genes, at least 330 genes, at least 335 genes, at least 340 genes, at least 345 genes, at least 350 genes, at least 355 genes, at least 360 genes, at least 365 genes, at least 370 genes, at least 375 genes, at least 380 genes, at least 385 genes, at least 390 genes, at least 395 genes, or at least 400 genes. In other embodiments, the genomic profile includes at least 325 genes. In certain embodiments, the genomic profile comprises at least 315 cancer-associated genes and introns in 28 genes (FOUNDATIONONE®) or the complete DNA coding sequence of 406 genes, introns in 31 rearranged genes, and RNA sequences (cDNA) of 265 genes (FOUNDATIONONE® Heme). In other embodiments, the genomic profile comprises 26 genes and 1000 associated mutations (EXODX® Solid Tumor). In yet other embodiments, the genomic profile comprises 76 genes (Guardant360). In yet other embodiments, the genomic profile comprises 73 genes (Guardant360). In other embodiments, the genomic profile comprises 354 genes and introns in 28 genes for rearrangement (FOUNDATIONONE® CDX TM In one embodiment, the genomic profile is FOUNDATIONONE® F1CDx. In another embodiment, the genomic profile comprises 468 genes (MSK-IMPACT TM). One or more genes may be added to the genomic profile as more genes are identified as being associated with oncology.

[0173] FOUNDATIONONE® Assay The FOUNDATIONONE® assay is a comprehensive genomic profiling assay for solid tumors, including, but not limited to, lung, colon, and breast solid tumors, melanoma, and ovarian cancer. The FOUNDATIONONE® assay uses hybrid capture, next-generation sequencing testing to identify genomic alterations (base substitutions, insertions and deletions, copy number alterations, and rearrangements), and genomic signature (e.g., TMB and microsatellite instability) selection. The assay covers 322 unique genes, including the entire coding regions of 315 cancer-related genes and selected introns from 28 genes. A complete list of FOUNDATIONONE® assay genes is provided in Tables 2 and 3. See FOUNDATIONONE: Technical Specifications, Foundation Medicine, Inc., available at FoundationMedicine.com (last accessed March 16, 2018), which is incorporated herein by reference in its entirety. [Table 2] [Table 3]

[0174] EXODX® Solid Tumor Assay In one embodiment, TMB is measured using the EXODX® Solid Tumor Assay. The EXODX® Solid Tumor Assay is an exoRNA and cfDNA-based assay that detects actionable mutations in cancer pathways. The EXODX® Solid Tumor Assay is a plasma-based assay that does not require tissue samples. The EXODX® Solid Tumor Assay covers 26 genes and 1,000 mutations. The specific genes covered by the EXODX® Solid Tumor Assay are listed in Table 4. See Plasma-Based Solid Tumor Mutation Panel Liquid Biopsy, Exosome Diagnostics, Inc., available at exosomedx.com (last accessed March 16, 2018). [Table 4]

[0175] FOUNDATIONONE® Liquid Assay In one embodiment, TMB is measured using the FOUNDATIONONE® Liquid Assay. The FOUNDATIONONE® Liquid Assay is a cfDNA-based assay that detects circulating tumor DNA (ctDNA). The assay is a plasma-based assay that does not require a solid tissue sample. The FOUNDATIONONE® Liquid Assay covers 70 genes. The specific genes covered by the FOUNDATIONONE® Liquid Assay are listed in Tables 5A-5C. See FOUNDATIONONE® Liquid, Technical Specifications, Foundation Medicine, available at assets.ctfassets.net / vhribv12lmne / 3SPYAcbGdqAeMsOqMyKUog / d0eb51659e08d733bf39971e85ed940d / F1L_TechnicalInformation_MKT-0061-04.pdf (last accessed October 6, 2018). [Table 5A] [Table 5B] [Table 5C]

[0176] Guardant360 assay In some embodiments, TMB status is determined using the Guardant360 assay. The Guardant360 assay measures mutations in at least 73 genes (Table 6), 23 indels (Table 7), 18 CNVs (Table 8), and 6 fusion genes (Table 9). See GuardantHealth.com (last accessed March 16, 2018). In some embodiments, TMB status is measured using the GUARDANTOMNI assay. TM Determined using an assay. TM The assay is a comprehensive genomic profiling tool that includes a 500-gene panel. [Table 6] [Table 7] [Table 8] [Table 9]

[0177] ILLUMINA® TruSight Assay In one embodiment, TMB is determined using the TruSight Tumor 170 Assay (ILLUMINA). The TruSight Tumor 170 Assay is a next-generation sequencing assay that simultaneously analyzes DNA and RNA and covers 170 genes associated with common solid tumors. The TruSight Tumor 170 Assay evaluates fusions, splice variants, insertions / deletions, single nucleotide variants (SNVs), and amplifications. A list of TruSight Tumor 170 Assay genes is shown in Tables 10-12. [Table 10] [Table 11] [Table 12]

[0178] FOUNDATIONONE® F1CDx Assay FOUNDATIONONE® CDX TM ("F1CDx") is a next-generation sequencing-based in vitro diagnostic device for the detection of genomic signatures, including substitutions, insertion and deletion alterations (indels), and copy number alterations (CNAs) in 324 genes and select gene rearrangements, as well as microsatellite instability (MSI) and tumor mutation burden (TMB), using DNA isolated from formalin-fixed, paraffin-embedded (FFPE) tumor tissue specimens. F1CDx has been approved by the U.S. Food and Drug Administration (FDA) for several tumor indications, including NSCLC, melanoma, breast cancer, colorectal cancer, and ovarian cancer.

[0179] The F1CDx assay uses a single DNA extraction method from routine FFPE biopsies or surgical resection specimens, with 50–1000 ng of DNA subjected to whole-genome shotgun library construction and hybridization-based capture of all coding exons from 309 cancer-related genes, one promoter region from 34 commonly rearranged genes (21 of which are also coding exons), one non-coding (ncRNA) region, and selected intron regions. Tables 13A and 13B provide a complete list of genes included in the F1CDx assay. In total, the assay detects alterations in 324 genes. The ILLUMINA® HiSeq 4000 platform is used to sequence the hybrid capture-selection libraries to high and uniform depth (>99% of exons with a target median coverage of >500× and coverage of >100×). The sequence data are then processed using a customized analysis pipeline designed to detect all classes of genomic alterations, including base substitutions, indels, copy number alterations (amplifications and homozygous gene deletions), and selected genomic rearrangements (e.g., gene fusions). Additionally, genomic signatures, including microsatellite instability (MSI) and tumor mutation burden (TMB), are reported. [Table 13A] [Table 13B]

[0180] The F1CDx assay identifies a variety of alterations in gene and / or intron sequences, including substitutions, insertions / deletions, and CNAs. The F1CDx assay was previously identified as having concordance with externally validated NGS assays and the FOUNDATIONONE® (F1 LDT) assay. FOUNDATIONONE® CDX, available at FoundationMedicine.com (last accessed March 16, 2018), is incorporated herein by reference in its entirety. TMSee: Technical Information, Foundation Medicine, Inc.

[0181] MSK-IMPACT TM In one embodiment, TMB status is measured using MSK-IMPACT TM Assay: MSK-IMPACT TM The assay uses next-generation sequencing to analyze the mutation status of 468 genes. Target genes are identified using the ILLUMINA HISEQ TM Captured and sequenced by the device. MSK-IMPACT TM The assay is USFDA approved for the detection of somatic mutations and microsatellite instability in solid malignant neoplasms. TM A complete list of the 468 genes analyzed by the assay is shown in Table 14. See Evaluation of Automatic Class III Designation for MSK-IMPACT (Integrated Mutation Profiling of Actionable Cancer Targets): Decision Summary, United States Food and Drug Administration, November 15, 2017, available at accessdata.fda.gov. [Table 14-1] [Table 14-2]

[0182] NEOGENOMICS(R) NEOTYPE TM Assay In some embodiments, TMB is administered as NEOGENOMICS® NEOTYPETM In one embodiment, TMB is determined using a NEOTYPE assay. TM In one embodiment, TMB is determined using the NEOTYPE solid tumor profile. The NEOGENOMICS assay measures the number of nonsynonymous DNA coding sequence changes per megabase pair of sequenced DNA.

[0183] ONCOMINE TM Tumor mutation burden assay In some embodiments, TMB is administered in a manner consistent with the use of THERMOFISHER SCIENTIFIC® ONCOMINE TM In one embodiment, TMB is determined using a tumor mutation assay. TM ONCOMINE TM Determined using a tumor mutation assay. TM ONCOMINE TM The Tumor Mutation Assay is a targeted NGS assay that quantifies somatic mutations to determine tumor mutation burden. The assay covers 1.7 Mb of DNA.

[0184] NOVOGENE TM NOVOPM TM Assay In one embodiment, TMB is administered by administration of NOVOGENE TM NOVOPM TM In one embodiment, TMB is determined using an assay. TM NOVOPM TM Determined using a cancer panel assay. TM NOVOPM TMThe Cancer Panel Assay is a comprehensive NGS cancer panel that analyzes the complete coding regions of 548 genes and introns of 21 genes, representing approximately 1.5 Mb of DNA, and is relevant to the diagnosis and / or treatment of solid tumors according to National Comprehensive Cancer Network (NCCN) guidelines and medical literature. The assay detects SNVs, InDels, fusions, and copy number variations (CNVs) genomic aberrations.

[0185] Other TMB assays In certain embodiments, TMB is determined using a TMB assay provided by CARIS® Life Sciences. In certain embodiments, TMB is determined using the PESONALIS® ACE ImmunoID assay. In certain embodiments, TMB is measured using PGDX® CANCERXOME TM Determined using the -R assay.

[0186] In yet other specific embodiments, genomic profiling detects all variants, i.e., single nucleotide variants, insertions / deletions (indels), copy number variations and rearrangements, e.g., translocations, expression and epigenetic markers.

[0187] A comprehensive gene panel often includes predetermined genes selected based on the type of tumor being analyzed.Therefore, the genomic profile used to measure TMB status can be selected based on the type of tumor that the subject has.In some embodiments, the genomic profile can include a set of genes specific to solid tumors.In other embodiments, the genomic profile can include a set of genes specific to hematological tumors and sarcomas.

[0188] ABL1, BRAF, CHEK1, FANCC, GATA3, JAK2, MITF, PD CD1LG2, RBM10, STAT4, ABL2, BRCA1, CHEK2, FANCD2, GATA4, JAK3, MLH1, PDGF RA, RET, STK11, ACVR1B, BRCA2, CIC, FANCE, GATA6, JUN, MPL, PDGFRB, RICTO R, SUFU, AKT1, BRD4, CREBBP, FANCF, GID4(C17orf39), KAT6A(MYST3), MRE11 A, PDK1, RNF43, SYK, AKT2, BRIP1, CRKL, FANCG, GLI1, KDM5A, MSH2, PIK3C2B ROS1, TAF1, AKT3, BTG1, CRLF2, FANCL, GNA11, KDM5C, MSH6, PIK3CA, RPTOR. TBX3, ALK, BTK, CSF1R, FAS, GNA13, KDM6A, MTOR, PIK3CB, RUNX1, TERC, AMER 1(FAM123B), C11orf30(EMSY), CTCF, FAT1, GNAQ, KDR, MUTYH, PIK3CG, RUNX1 T1, TERT(Type), APC, CARD11, CTNNA1, FBXW7, GNAS, KEAP1, MYC, PIK3R 1, SDHA, TET2, AR, CBFB, CTNNB1, FGF10, GPR124, KEL, MYCL(MYCL1), PIK3R2. SDHB, TGFBR2, ARAF, CBL, CUL3, FGF14, GRIN2A, KIT, MYCN, PLCG2, SDHC, and TNF AIP3, ARFRP1, CCND1, CYLD, FGF19, GRM3, KLHL6, MYD88, PMS2, SDHD, TNFRSF1 4. ARID1A, CCND2, DAXX, FGF23, GSK3B, KMT2A(MLL), NF1, POLD1, SETD2, TOP 1, ARID1B, CCND3, DDR2, FGF3, H3F3A, KMT2C(MLL3), NF2, POLE, SF3B1, and TOP2A ARID2, CCNE1, DICER1, FGF4, HGF, KMT2D(MLL2), NFE2L2, PPP2R1A, SLIT2, T P53, ASXL1, CD274, DNMT3A, FGF6, HNF1A, KRAS, NFKBIA, PRDM1, SMAD2, TSC1.ATM, CD79A, DOT1L, FGFR1, HRAS, LMO1, NKX2-1, PREX2, SMAD3, TSC2, ATR, CD79B, EGFR, FGFR2, HSD3B1, LRP1B, NOTCH1, PRKAR1A, SMAD4, TSHR , ATRX, CDC73, EP300, FGFR3, HSP90AA1, LYN, NOTCH2, PRKCI, SMARCA4, U2AF1, AURKA, CDH1, EPHA3, FGFR4, IDH1, LZTR1, NOTCH3, PRKDC, SMARC B1, VEGFA, AURKB, CDK12, EPHA5, FH, IDH2, MAGI2, NPM1, PRSS8, SMO, VHL, AXIN1, CDK4, EPHA7, FLCN, IGF1R, MAP2K1, NRAS, PTCH1, SNCAIP, WI SP3, AXL, CDK6, EPHB1, FLT1, IGF2, MAP2K2, NSD1, PTEN, SOCS1, WT1, BAP1, CDK8, ERBB2, FLT3, IKBKE, MAP2K4, NTRK1, PTPN11, SOX10, XPO1, BA RD1, CDKN1A, ERBB3, FLT4, IKZF1, MAP3K1, NTRK2, QKI, SOX2, ZBTB2, BCL2, CDKN1B, ERBB4, FOXL2, IL7R, MCL1, NTRK3, RAC1, SOX9, ZNF217, BC L2L1, CDKN2A, ERG, FOXP1, INHBA, MDM2, NUP93, RAD50, SPEN, ZNF703, BCL2L2, CDKN2B, ERRFI1, FRS2, INPP4B, MDM4, PAK3, RAD51, SPOP, BCL6, In other embodiments, the TMB analysis further comprises one or more genes selected from the group consisting of CDKN2C, ESR1, FUBP1, IRF2, MED12, PALB2, RAF1, SPTA1, BCOR, CEBPA, EZH2, GABRA6, IRF4, MEF2B, PARK2, RANBP2, SRC, BCORL1, CHD2, FAM46C, GATA1, IRS2, MEN1, PAX5, RARA, STAG2, BLM, CHD4, FANCA, GATA2, JAK1, MET, PBRM1, RB1, STAT3, and any combination thereof.This includes identifying genomic alterations in one or more of ETV6 and MYB.

[0189] These are ABL1, 12B, ABL2, ACTB, ACVR1, and ACVR 1B, AGO2, AKT1, AKT2, AKT3, ALK, ALOX, ALOX12B, AMER1, AMER1 (FAM123BまたはWTX)、AMER1(FAM123B)、ANKRD11、APC、APH1A、AR、ARAF、ARFRP1、ARHGAP26(GRAF)、ARID1A、ARID1B、ARID2、YES D5B, ARv7, ASMTL, ASXL1, ASXL2, ATM, ATR, ATRX, AURKA, AURKB AXIN1, AXIN2, AXL, B2M, BABAM1, BAP1, BARD1, BBC3, BCL10, B CL11B, BCL2, BCL2L1, BCL2L11, BCL2L2, BCL6, BCL7A, BCOR, BCORL1, BIRC3, BLM, BMPR1A, BRAF, BRCA1, BRCA2, BRD4, BRIP1, B RIP1(BACH1), BRSK1, BTG1, BTG2, BTK, BTLA, C11orf30(EMSY), C11orf30, C11orf30(EMSY), CAD, CALR, CARD11, CARM1, CASP 8. CBFB, CBL, CCND1, CCND2, CCND3, CCNE1, CCT6B, CD22, CD274, CD274(PD-L1), CD276, CD36, CD58, CD70, CD79A, CD79B, CDC4 2. CDC73, CDH1, CDK12, CDK4, CDK6, CDK8, CDKN1A, CDKN1B, CDKN2A, CDKN2Ap14ARF, CDKN2Ap16INK4A, CDKN2B, CDKN2C, CEBPA CENPA, CHD2, CHD4, CHEK1, CHEK2, CIC, CIITA, CKS1B, CPS1, CREBBP, CRKL, CRLF2, CSDE1, CSF1R, CSF3R, CTCF, CTLA-4, CTNN B1, CTNNA1, CTNNB1, CUL3, CUL4A, CUX1, CXCR4, CYLD, CYP17A1, CYSLTR2, DAXX, DCUN1D1, DDR1, DDR2, DDX3X, DH 2, DICER1, DIS3, DNAJB1, DNM2, DNMT1, DNMT3A, DNMT3B, DOT1L, DROSHA, DTX1, DUSP2, DUSP4, DUSP9, E2F3, EBF1.ECT2L, EED, EGFL7, EGFR, EIF1AX, EIF4A2, EIF4E, ELF3, ELP2, EML4, EML4-ALK, EP300, EPAS 1. EPCAM, EPHA3, EPHA5, EPHA7, EPHB1, EPHB4, ERBB2, ERBB3, ERBB4, ERCC1, ERCC2, ERCC3, E RCC4, ERCC5, ERF, ERG, ERRFI1, ERRFl1, ESR1, ETS1, ETV1, ETV4, ETV5, ETV6, EWSR1, EXOSC6 EZH1, EZH2, FAF1, FAM175A, FAM46C, FAM58A, FANCA, FANCC, FANCD2, FANCE, FANCF, FANCG FANCI, FANCL, FAS, FAS(TNFRSF6), FAT1, FBXO11, FBXO31, FBXW7, FGF1, FGF10, FGF12, FGF1 4. FGF19, FGF2, FGF23, FGF3, FGF4, FGF5, FGF6, FGF7, FGF8, FGF9, FGFR1, FGFR2, FGFR3, FGF R4, FH, FHIT, FLCN, FLI1, FLT1, FLT3, FLT4, FLYWCH1, FOXA1, FOXL2, FOXO1, FOXO3, FOXP1, F RS2、FUBP1、FYN、GABRA6、GADD45B、GATA1、GATA2、GATA3、GATA4、GATA6、GEN1、GID4(C17orf 39) GID4(C17orf39), GLI1, GLl1, GNA11, GNA12, GNA13, GNAQ, GNAS, GPR124, GPS2, GREM1, GRIN2A, GRM3, GSK3B, GTSE1 H3F3A, H3F3B, H3F3C, HDAC1, HDAC4, HDAC7, HER-2 / NEU;ERBB2、HGF、HIST1H1C、HIST1H1D、HIST1H1E、HIST1H2A C、HIST1H2AG、HIST1H2AL、HIST1H2AM、HIST1H2BC、HIST1H2BD、HIST1H2BJ、HIST1H2BK、HIST1H2BO、HIST1H3A、HIST1H3B HIST1H3C、HIST1H3D、HIST1H3E、HIST1H3F、HIST1H3G、HIST1H3H、HIST1H3I、HIST1H3J、HIST2H3C、HIST2H3D、HIST3H3HLA-A, HLA-B, HNF1A, HOXB13, HRAS, HSD3B1, HSP90AA1, IK, ICOSLG, ID3, IDH1, IDH2, IFNGR1, IGF1, IGF1R, IGF2, IKBKE, IKZF1, IKZF2, IKZF3, IL10, I L7R、INHA、INHBA、INPP4A、INPP4B、INPP5D(SHIP)、INPPL1、INSR、IRF1、IRF 2、IRF4、IRF8、IRS1、IRS2、JAK1、JAK2、JAK3、JARID2、JUN、K14、KAT6A(MYST) 3) KAT6A(MYST3), KDM2B, KDM4C, KDM5A, KDM5C, KDM6A, KDR, KEAP1, KEL, KIF5B, KIT, KLF4, KLHL6, KMT2A, KMT2A(MLL), KMT2B, KMT2C, KM T2C(MLL3)、KMT2D、KMT2D(MLL2)、KNSTRN、KRAS、LAMP1、LATS1、LATS 2、LEF1、LMO1、LRP1B、LRRK2、LTK、LYN、LZTR1、MAF、MAFB、MAGED1、MA GI2、MALT1、MAP2K1、MAP2K1(MEK1)、MAP2K2、MAP2K2(MEK2)、MAP2K4、MAP3、MAP3K1、MAP3K13、MAP3K14、MAP3K6、MAP3K7、MAPK1、MAPK3、M APKAP1、MAX、MCL1、MDC1、MDM2、MDM4、MED12、MEF2B、MEF2C、MEK1、ME N1、MERTK、MET、MGA、MIB1、MITF、MKI67、MKNK1、MLH1、MLLT3、MPL、MRE 11A、MRE11A、MSH2、MSH3、MSH6、MSI1、MSI2、MST1、MST1R、MTAP、MTOR、MUTYH、MYC、MYCL、MYCL(MYC) L1)、MYCL(MYCL1)、MYCL1、MYCN、MYD88、MYO18A、MYOD1、NBN、NCOA 3、NCOR1、NCOR2、NCSTN、NEGR1、NF1、NF2、NFE2L2、NFKBIA、NKX2-1 、NKX3-1、NOD1、NOTCH1、NOTCH2、NOTCH3、NOTCH4、NPM1、NRAS、NRG 1、NSD1、NT5C2、NTHL1、NTRK1、NTRK2、NTRK3、NUF2、NUP93、NUP98、P2RY8、PAG1、PAK1、PAK3、PAK7、PALB2、PARK2、PARP1、PARP2、PARP3、PASK、PAX3、PAX5、PAX7、PBRM1、PC、PCBP1、PCLO、PDCD1、PDCD1(PD-1)、PDCD11、PDCD1LG2、PDCD1LG2(PD-L2)、PDGFRA、PDGFRB、PDK1、PDPK1、PGR、PHF6、PHOX2B、PIK3C2B、PIK3C2G、PIK3C3、PIK3CA、PIK3CB、PIK3CD、PIK3CG、PIK3R1、PIK3R2、PIK3R3、PIM1、PLCG2、PLK2、PMAIP1、PMS1、PMS2、PNRC1、POLD1、POLE、POT1、PPARG、PPM1D、PPP2、PPP2R1A、PPP2R2A、PPP4R2、PPP6C、PRDM1、PRDM14、PREX2、PRKAR1A、PRKCI、PRKD1、PRKDC、PRSS8、PTCH1、PTEN、PTP4A1、PTPN11、PTPN2、PTPN6(SHP-1)、PTPRD、PTPRO、PTPRS、PTPRT、QKI、R1A、RAB35、RAC1、RAC2、RAD21、RAD50、RAD51、RAD51B、RAD51C、RAD51D、RAD52、RAD54L、RAF1、RANBP2、RARA、RASA1、RASGEF1A、RB1、RBM10、RECQL、RECQL4、REL、RELN、RET、RFWD2、RHEB、RHOA、RICTOR、RIT1、RNF43、ROS1、RPS6KA4、RPS6KB1、RPS6KB2、RPTOR、RRAGC、RRAS、RRAS2、RTEL1、RUNX1、RUNX1T1、RXRA、RYBP、S1PR2、SDHA、SDHAF2、SDHB、SDHC、SDHD、SERP2、SESN1、SESN2、SESN3、SETBP1、SETD2、SETD8、SF3B1、SGK1、SH2B3、SH2D1A、SHOC2、SHQ1、SLIT2、SLX4、SMAD2、SMAD3、SMAD4、SMARCA1、SMARCA4、SMARCB1、SMARCD1、SMC1A、SMC3、SMO、SMYD3、SNCAIP、SOCS1、SOCS2、SOCS3、SOS1、SOX10、SOX17、SOX2、SOX9、SPEN、SPOP、SPRED1、SPTA1, SRC, SRSF2, STAG2, STAT3, STAT4, STAT5A, STAT5B, STAT6, STK11, STK19, STK40, SUFU, SUZ12, SYK, TAF1, TAP1, TAP2, TBL1XR1, TBX3, TCEB1, TCF3, TCF3(E2A), TCF7L2, TCL1A( TCL1), TEK, TERC, TERT, TERT promoter, TET1, TET2, TFRC, TGFBR1, TGFBR2, TIPARP, TLL2, TMEM127, TMEM30A, TMPRSS2, TMSB4XP8(TMSL3), TNFAIP3, TNFRSF11A, TNFRSF14, TNFRSF17, TO The gene includes one or more genes selected from the group consisting of P1, TOP2A, TP53, TP53BP1, TP63, TRAF2, TRAF3, TRAF5, TRAF7, TSC1, TSC2, TSHR, TUSC3, TYK2, TYRO3, U2AF1, U2AF2, UPF1, VEGFA, VHL, VTCN1, WDR90, WHSC1, WHSC1 (MMSET or NSD2), WHSC1L1, WISP3, WT1, WWTR1, XBP1, XIAP, XPO1, XRCC2, YAP1, YES1, YY1AP1, ZBTB2, ZFHX3, ZMYM3, ZNF217, ZNF24 (ZSCAN3), ZNF703, ZRSR2, and any combination thereof.

[0190] In other embodiments, the genomic profiling assay is performed using a gene encoding ABL1, 12B, ABL2, ACTB, ACVR1, ACVR1B, AGO2, AKT1, AKT2, AKT3, ALK, ALOX, ALOX12B, AMER1, AMER1 (FAM123B or WTX), AMER1 (FAM123B), ANKRD11, APC, APH1A, AR, ARAF, ARFRP1, ARHGAP26 (GRAF), ARID1A, ARID1B, ARID 2, ARID5B, ARv7, ASMTL, ASXL1, ASXL2, ATM, ATR, ATRX, AURKA, AURKB, AXIN1, AXIN2, AXL, B2M, BABAM1, BAP1, BARD1, BBC3, BCL 10, BCL11B, BCL2, BCL2L1, BCL2L11, BCL2L2, BCL6, BCL7A, BCOR, BCORL1, BIRC3, BLM, BMPR1A, BRAF, BRCA1, BRCA2, BRD4, BRIP 1, BRIP1(BACH1), BRSK1, BTG1, BTG2, BTK, BTLA, C11orf30(EMSY), C11orf30, C11orf30(EMSY), CAD, CALR, CARD11, CARM1, C ASP8, CBFB, CBL, CCND1, CCND2, CCND3, CCNE1, CCT6B, CD22, CD274, CD274(PD-L1), CD276, CD36, CD58, CD70, CD79A, CD79B, CD C42, CDC73, CDH1, CDK12, CDK4, CDK6, CDK8, CDKN1A, CDKN1B, CDKN2A, CDKN2Ap14ARF, CDKN2Ap16INK4A, CDKN2B, CDKN2C, CEBP A, CENPA, CHD2, CHD4, CHEK1, CHEK2, CIC, CIITA, CKS1B, CPS1, CREBBP, CRKL, CRLF2, CSDE1, CSF1R, CSF3R, CTCF, CTLA-4, CTNN B1, CTNNA1, CTNNB1, CUL3, CUL4A, CUX1, CXCR4, CYLD, CYP17A1, CYSLTR2, DAXX, DCUN1D1, DDR1, DDR2, DDX3X, DH2, DICER1, DIS3, DNAJB1, DNM2, DNMT1, DNMT3A, DNMT3B, DOT1L, DROSHA, DTX1, DUSP2, DUSP4, DUSP9, E2F3,EBF1, ECT2L, EED, EGFL7, EGFR, EIF1AX, EIF4A2, EIF4E, ELF3, ELP2, EML4, EML4-ALK, EP300 EPAS1, EPCAM, EPHA3, EPHA5, EPHA7, EPHB1, EPHB4, ERBB2, ERBB3, ERBB4, ERCC1, ERCC2, ERC C3、ERCC4、ERCC5、ERF、ERG、ERRFI1、ERRFl1、ESR1、ETS1、ETV1、ETV4、ETV5、ETV6、EWSR1、EX OSC6, EZH1, EZH2, FAF1, FAM175A, FAM46C, FAM58A, FANCA, FANCC, FANCD2, FANCE, FANCF, FAN CG, FANCI, FANCL, FAS, FAS(TNFRSF6), FAT1, FBXO11, FBXO31, FBXW7, FGF1, FGF10, FGF12, F GF14, FGF19, FGF2, FGF23, FGF3, FGF4, FGF5, FGF6, FGF7, FGF8, FGF9, FGFR1, FGFR2, FGFR3 GFR4, FH, FHIT, FLCN, FLI1, FLT1, FLT3, FLT4, FLYWCH1, FOXA1, FOXL2, FOXO1, FOXO3, FOXP1 FRS2、FUBP1、FYN、GABRA6、GADD45B、GATA1、GATA2、GATA3、GATA4、GATA6、GEN1、GID4(C17orf 39) GID4(C17orf39), GLI1, GLl1, GNA11, GNA12, GNA13, GNAQ, GNAS, GPR124, GPS2, GREM1, GRIN2A, GRM3, GSK3B, GTSE1 H3F3A, H3F3B, H3F3C, HDAC1, HDAC4, HDAC7, HER-2 / NEU;ERBB2、HGF、HIST1H1C、HIST1H1D、HIST1H1E、HIST1H2A C、HIST1H2AG、HIST1H2AL、HIST1H2AM、HIST1H2BC、HIST1H2BD、HIST1H2BJ、HIST1H2BK、HIST1H2BO、HIST1H3A、HIST1H3B HIST1H3C、HIST1H3D、HIST1H3E、HIST1H3F、HIST1H3G、HIST1H3H、HIST1H3I、HIST1H3J、HIST2H3C、HIST2H3D、HIST3H3HLA-A, HLA-B, HNF1A, HOXB13, HRAS, HSD3B1, HSP90AA1, IK, ICOSLG, ID3, IDH1, IDH2, IFNGR1, IGF1, IGF1R, IGF2, IKBKE, IKZF1, IKZF2, IKZF3, IL10, I L7R、INHA、INHBA、INPP4A、INPP4B、INPP5D(SHIP)、INPPL1、INSR、IRF1、IRF 2、IRF4、IRF8、IRS1、IRS2、JAK1、JAK2、JAK3、JARID2、JUN、K14、KAT6A(MYST) 3) KAT6A(MYST3), KDM2B, KDM4C, KDM5A, KDM5C, KDM6A, KDR, KEAP1, KEL, KIF5B, KIT, KLF4, KLHL6, KMT2A, KMT2A(MLL), KMT2B, KMT2C, KM T2C(MLL3)、KMT2D、KMT2D(MLL2)、KNSTRN、KRAS、LAMP1、LATS1、LATS 2、LEF1、LMO1、LRP1B、LRRK2、LTK、LYN、LZTR1、MAF、MAFB、MAGED1、MA GI2、MALT1、MAP2K1、MAP2K1(MEK1)、MAP2K2、MAP2K2(MEK2)、MAP2K4、MAP3、MAP3K1、MAP3K13、MAP3K14、MAP3K6、MAP3K7、MAPK1、MAPK3、M APKAP1、MAX、MCL1、MDC1、MDM2、MDM4、MED12、MEF2B、MEF2C、MEK1、ME N1、MERTK、MET、MGA、MIB1、MITF、MKI67、MKNK1、MLH1、MLLT3、MPL、MRE 11A、MRE11A、MSH2、MSH3、MSH6、MSI1、MSI2、MST1、MST1R、MTAP、MTOR、MUTYH、MYC、MYCL、MYCL(MYC) L1)、MYCL(MYCL1)、MYCL1、MYCN、MYD88、MYO18A、MYOD1、NBN、NCOA 3、NCOR1、NCOR2、NCSTN、NEGR1、NF1、NF2、NFE2L2、NFKBIA、NKX2-1 、NKX3-1、NOD1、NOTCH1、NOTCH2、NOTCH3、NOTCH4、NPM1、NRAS、NRG 1、NSD1、NT5C2、NTHL1、NTRK1、NTRK2、NTRK3、NUF2、NUP93、NUP98、P2RY8、PAG1、PAK1、PAK3、PAK7、PALB2、PARK2、PARP1、PARP2、PARP3、PASK、PAX3、PAX5、PAX7、PBRM1、PC、PCBP1、PCLO、PDCD1、PDCD1(PD-1)、PDCD11、PDCD1LG2、PDCD1LG2(PD-L2)、PDGFRA、PDGFRB、PDK1、PDPK1、PGR、PHF6、PHOX2B、PIK3C2B、PIK3C2G、PIK3C3、PIK3CA、PIK3CB、PIK3CD、PIK3CG、PIK3R1、PIK3R2、PIK3R3、PIM1、PLCG2、PLK2、PMAIP1、PMS1、PMS2、PNRC1、POLD1、POLE、POT1、PPARG、PPM1D、PPP2、PPP2R1A、PPP2R2A、PPP4R2、PPP6C、PRDM1、PRDM14、PREX2、PRKAR1A、PRKCI、PRKD1、PRKDC、PRSS8、PTCH1、PTEN、PTP4A1、PTPN11、PTPN2、PTPN6(SHP-1)、PTPRD、PTPRO、PTPRS、PTPRT、QKI、R1A、RAB35、RAC1、RAC2、RAD21、RAD50、RAD51、RAD51B、RAD51C、RAD51D、RAD52、RAD54L、RAF1、RANBP2、RARA、RASA1、RASGEF1A、RB1、RBM10、RECQL、RECQL4、REL、RELN、RET、RFWD2、RHEB、RHOA、RICTOR、RIT1、RNF43、ROS1、RPS6KA4、RPS6KB1、RPS6KB2、RPTOR、RRAGC、RRAS、RRAS2、RTEL1、RUNX1、RUNX1T1、RXRA、RYBP、S1PR2、SDHA、SDHAF2、SDHB、SDHC、SDHD、SERP2、SESN1、SESN2、SESN3、SETBP1、SETD2、SETD8、SF3B1、SGK1、SH2B3、SH2D1A、SHOC2、SHQ1、SLIT2、SLX4、SMAD2、SMAD3、SMAD4、SMARCA1、SMARCA4、SMARCB1、SMARCD1、SMC1A、SMC3、SMO、SMYD3、SNCAIP、SOCS1、SOCS2、SOCS3、SOS1、SOX10、SOX17、SOX2、SOX9、SPEN、SPOP、SPRED1、SPTA1, SRC, SRSF2, STAG2, STAT3, STAT4, STAT5A, STAT5B, STAT6, STK11, STK19, STK40, SUFU, SUZ12, SYK, TAF1, TAP1, TAP2, TBL1XR1 , TBX3, TCEB1, TCF3, TCF3(E2A), TCF7L2, TCL1A(TCL1), TEK, TERC, TERT, TERT promoter, TET1, TET2, TFRC, TGFBR1, TGFBR2, TIPARP, TLL 2, TMEM127, TMEM30A, TMPRSS2, TMSB4XP8 (TMSL3), TNFAIP3, TNFRSF11A, TNFRSF14, TNFRSF17, TOP1, TOP2A, TP53, TP53BP1, TP63, TRAF2, TRAF3, TRAF5, TRAF7, TSC1, TSC2, TSHR, TUSC3, TYK2, TYRO3, U2AF1, U2AF2, UPF1, VEGFA, VHL, VTCN1, WDR90, WHSC1, WHSC1 (MMSET or or NSD2), WHSC1L1, WISP3, WT1, WWTR1, XBP1, XIAP, XPO1, XRCC2, YAP1, YES1, YY1AP1, ZBTB2, ZFHX3, ZMYM3, ZNF217, ZNF24 (ZSCAN3), ZNF703, ZRSR2, and any combination thereof, 0, at least about 100, at least about 110, at least about 120, at least about 130, at least about 140, at least about 150, at least about 160, at least about 170, at least about 180, at least about 190, at least about 200, at least about 210, at least about 220, at least about 230, at least about 240, at least about 250, at least about 260, at least about 270, at least about 280, at least about 290 or at least about 300 genes.

[0191] In other embodiments, the genomic profile comprises one or more genes selected from the genes listed in Tables 2-14.

[0192] In some embodiments, the TMB status based on genomic profiling is highly correlated with the TMB status based on whole exome or whole genome sequencing.There is evidence that the use of genomic profiling assays such as F1CDx assays has the same performance as whole exome and / or whole genome sequencing assays.These data support the use of genomic profiling assays as a more efficient means of measuring TMB status without losing the prognostic quality of TMB status.

[0193] TMB can be measured using tissue biopsy samples, or alternatively, circulating tumor DNA (ctDNA), cfDNA (cell-free DNA), and / or liquid biopsy samples. ctDNA can be used to measure TMB status by whole-exome or whole-genome sequencing or genomic profiling using available methods, e.g., GRAIL, Inc.

[0194] Subjects with SCLC-derived tumors are identified as suitable for treatment with anti-PD-1 antibody monotherapy or anti-PD-1 / anti-CTLA-4 combination therapy based on measuring TMB status and identifying high TMB. In certain embodiments, the TMB score is calculated as the total number of nonsynonymous missense mutations in the tumor as measured by whole exome sequencing or whole genome sequencing. In certain embodiments, a high TMB is defined as a mutation count of at least 210, at least 215, at least 220, at least 225, at least 230, at least 235, at least 240, at least 245, at least 250, at least 255, at least 260, at least 265, at least 270, at least 275, at least 280, at least 285, at least 290, at least 295, at least 300, at least 305, at least 310, at least 315, at least 320, at least 325, at least 330, at least 335, at least 340, at least 345, at least 350, at least 360, at least 370, at least 380, at least 390, at least 400, at least 410, at least 420, at least 430, at least 440, at least 450, at least 460, at least 470, at least 480, at least 490, at least 500, at least 510, at least 520, at least 530, at least 540, at least 550, at least 560, at least 570, at least 580, at least 590, at least 600, at least 610, at least 620, at least 630, at least 640, In some embodiments, the marker has a score of at least 355, at least 360, at least 365, at least 370, at least 375, at least 380, at least 385, at least 390, at least 395, at least 400, at least 405, at least 410, at least 415, at least 420, at least 425, at least 430, at least 435, at least 440, at least 445, at least 450, at least 455, at least 460, at least 465, at least 470, at least 475, at least 480, at least 485, at least 490, at least 495, or at least 500.In other embodiments, high TMB has a score of at least 215, at least 220, at least 221, at least 222, at least 223, at least 224, at least 225, at least 226, at least 227, at least 228, at least 229, at least 230, at least 231, at least 232, at least 233, at least 234, at least 235, at least 236, at least 237, at least 238, at least 239, at least 240, at least 241, at least 242, at least 243, at least 244, at least 245, at least 246, at least 247, at least 248, at least 249, or at least 250. In certain embodiments, high TMB has a score of at least 243. In other embodiments, high TMB has a score of at least 244. In certain embodiments, high TMB has a score of at least 245. In other embodiments, high TMB has a score of at least 246. In other embodiments, high TMB has a score of at least 247. In other embodiments, high TMB has a score of at least 248. In other embodiments, high TMB has a score of at least 249. In other embodiments, high TMB has a score of at least 250. In other embodiments, high TMB has a score of any integer between 200 and 300 or higher. In other embodiments, high TMB has a score of any integer between 210 and 290 or higher. In other embodiments, high TMB has a score of any integer between 220 and 280 or higher. In other embodiments, high TMB has a score of any integer between 230 and 270 or higher. In other embodiments, high TMB has a score of any integer between 235 and 265 or higher.

[0195] Alternatively, high TMB can be a relative value rather than an absolute value. In some embodiments, the subject's TMB status is compared to a control TMB value. In some embodiments, the subject's TMB status is within the highest quantile of the control TMB value. In other embodiments, the subject's TMB status is within the highest tertile of the control TMB value.

[0196] In some embodiments, TMB status is expressed as the number of mutations per sample, cell, exome, or DNA length (e.g., Mb). In some embodiments, a tumor is in a high TMB status if it has at least about 50 mutations / tumor, at least about 55 mutations / tumor, at least about 60 mutations / tumor, at least about 65 mutations / tumor, at least about 70 mutations / tumor, at least about 75 mutations / tumor, at least about 80 mutations / tumor, at least about 85 mutations / tumor, at least about 90 mutations / tumor, at least about 95 mutations / tumor, at least about 100 mutations / tumor, at least about 105 mutations / tumor, at least about 110 mutations / tumor, at least about 115 mutations / tumor, or at least about 120 mutations / tumor. In some embodiments, a tumor is in a high TMB state if it has at least about 125 mutations / tumor, at least about 150 mutations / tumor, at least about 175 mutations / tumor, at least about 200 mutations / tumor, at least about 225 mutations / tumor, at least about 250 mutations / tumor, at least about 275 mutations / tumor, at least about 300 mutations / tumor, at least about 350 mutations / tumor, at least about 400 mutations / tumor, or at least about 500 mutations / tumor. In certain embodiments, a tumor is in a high TMB state if it has at least about 100 mutations / tumor.

[0197] In some embodiments, the tumor has been genomically sequenced, e.g., genomically sequenced by a TMB assay, e.g., FOUNDATIONONE® CDX TMA tumor has a high TMB status if it has at least about 5 mutations per megabase pair of genome sequenced by the assay (mutations / Mb), at least about 6 mutations / Mb, at least about 7 mutations / Mb, at least about 8 mutations / Mb, at least about 9 mutations / Mb, at least about 10 mutations / Mb, at least about 11 mutations / Mb, at least about 12 mutations / Mb, at least about 13 mutations / Mb, at least about 14 mutations / Mb, at least about 15 mutations / Mb, at least about 20 mutations / Mb, at least about 25 mutations / Mb, at least about 30 mutations / Mb, at least about 35 mutations / Mb, at least about 40 mutations / Mb, at least about 45 mutations / Mb, at least about 50 mutations / Mb, at least about 75 mutations / Mb, or at least about 100 mutations / Mb. In some embodiments, a tumor has a high TMB status if it has at least about 5 mutations / Mb. In some embodiments, a tumor has a high TMB status if it has at least about 10 mutations / Mb. In some embodiments, a tumor has a high TMB status if it has at least about 11 mutations / Mb. In some embodiments, a tumor has a high TMB status if it has at least about 12 mutations / Mb. In some embodiments, a tumor has a high TMB status if it has at least about 13 mutations / Mb. In some embodiments, a tumor has a high TMB status if it has at least about 14 mutations / Mb. In some embodiments, a tumor has a high TMB status if it has at least about 15 mutations / Mb.

[0198] Because mutations may differ by tumor type and in other respects (see Q4 and Q5), the values ​​associated with "TMB high" and "TMB low" may differ by tumor type.

[0199] PD-L1 status TMB status can be used alone or in combination with other factors as a predictor of tumor response to therapy, particularly treatment with cancer immunotherapeutics such as anti-PD-1 antibodies or anti-CTLA-4 antibodies. In one embodiment, tumor TMB status alone is used to identify patients with tumors likely to respond to treatment with anti-PD-1 antibody monotherapy or anti-PD-1 / anti-CTLA-4 antibody combination therapy. In other embodiments, PD-L1 status and TMB status are used to identify patients with tumors likely to respond to treatment with anti-PD-1 antibody monotherapy or anti-PD-1 / anti-CTLA-4 antibody combination therapy.

[0200] The PD-L1 status of a tumor in a subject can be measured prior to administering any of the compositions or using any of the methods disclosed herein. PD-L1 expression can be determined by any method known in the art.

[0201] To assess PD-L1 expression, in certain embodiments, a test tissue sample may be obtained from a patient in need of treatment. In other embodiments, assessing PD-L1 expression may be achieved without obtaining a test tissue sample. In certain embodiments, selecting an appropriate patient involves (i) providing a test tissue sample containing tumor cells and / or tumor-infiltrating inflammatory cells, optionally obtained from a patient with cancer of the tissue; and (ii) assessing the proportion of cells in the test tissue sample that express PD-L1 on their cell surface based on determining that the proportion of cells in the test tissue sample that express PD-L1 on their cell surface is higher than a predetermined threshold level.

[0202] However, it should be understood that in any of the methods involving measuring PD-L1 expression in a test tissue sample, the step involving preparing a test tissue sample from a patient is an optional step. Also, in certain embodiments, the "measuring" or "assessing" step of identifying or determining the number or percentage of cells in the test tissue sample that express PD-L1 on their surface is performed by a novel method for assaying PD-L1 expression, such as performing a reverse transcriptase polymerase chain reaction (RT-PCR) assay or an IHC assay. In certain other embodiments, no novel steps are included, and PD-L1 expression is assessed, for example, by review of clinical laboratory reports. In certain embodiments, the methods up to and including the steps of assessing PD-L1 expression provide intermediate results that can be provided to a physician or other healthcare professional for use in selecting appropriate candidates for anti-PD-1 antibodies or anti-PD-L1 antibody therapy. In certain embodiments, the step of providing the intermediate result is performed by a physician or someone acting under the direction of a physician. In other embodiments, these steps are performed by an independent person, such as an independent laboratory or laboratory technician.

[0203] In certain embodiments of any of the methods, the proportion of cells expressing PD-L1 is determined by performing an assay to determine the presence of PD-L1 RNA. In further embodiments, the presence of PD-L1 RNA is determined by RT-PCR, in situ hybridization, or RNase protection. In other embodiments, the proportion of cells expressing PD-L1 is determined by performing an assay to determine the presence of PD-L1 polypeptide. In further embodiments, the presence of PD-L1 polypeptide is determined by immunohistochemistry (IHC), enzyme-linked immunosorbent assay (ELISA), in vivo imaging, or flow cytometry. In certain embodiments, PD-L1 expression is assayed by IHC. In other embodiments of all of these methods, cell surface expression of PD-L1 is assayed using, for example, IHC or in vivo imaging.

[0204] Imaging techniques have provided important tools for cancer research and treatment. Recent developments in molecular imaging systems, including positron emission tomography (PET), single-photon emission computed tomography (SPECT), fluorescence reflectance imaging (FRI), fluorescence-mediated tomography (FMT), bioluminescence imaging (BLI), confocal laser scanning microscopy (LSCM), and multiphoton microscopy (MPM), may herald greater use of these techniques in cancer research. Some of these molecular imaging systems allow physicians not only to see where tumors are located in the body, but also to visualize the expression and activity of specific molecules, cells, and biological processes that affect tumor behavior and / or responsiveness to therapeutic agents (Condeelis and Weissleder, "In vivo imaging in cancer," Cold Spring Harb. Perspect. Biol. 2(12):a003848 (2010)). Antibody specificity combined with the sensitivity and resolution of PET makes immunoPET imaging particularly attractive for monitoring and assaying antigen expression in tissue samples (McCabe and Wu, "Positive progress in immunoPET—not just a coincidence," Cancer Biother. Radiopharm. 25(3):253-61 (2010); Olafsen et al., "ImmunoPET imaging of B-cell lymphoma using 124I-anti-CD20 scFv dimers (diabodies)," Protein Eng. Des. Sel. 23(4):243-9 (2010)). In certain embodiments of any of the methods, PD-L1 expression is assayed by immunoPET imaging. In certain embodiments of any of the methods, the proportion of cells in the test tissue sample that express PD-L1 is assessed by performing an assay that determines the presence of PD-L1 polypeptide on the surface of cells in the test tissue sample. In certain embodiments, the test tissue sample is an FFPE tissue sample. In other embodiments, the presence of a PD-L1 polypeptide is determined by an IHC assay.In a further embodiment, the IHC assay is performed using automated processing. In one embodiment, the IHC assay is performed using an anti-PD-L1 monoclonal antibody that binds to a PD-L1 polypeptide. In one embodiment, the anti-PD-L1 monoclonal antibody is selected from the group consisting of 28-8, 28-1, 28-12, 29-8, 5H1, and any combination thereof. See WO / 2013 / 173223, the entire contents of which are incorporated herein by reference.

[0205] In one embodiment of this method, an automated IHC method is used to assay PD-L1 expression on the cell surface of an FFPE tissue specimen. The presence of human PD-L1 antigen can be measured in a test tissue sample by contacting the test sample and a negative control sample (e.g., normal tissue) with a monoclonal antibody that specifically binds to human PD-L1 under conditions that allow the antibody or a portion thereof to form a complex with human PD-L1. In one embodiment, the test and control tissue samples are FFPE samples. The formation of the complex is then detected, where a difference in complex formation between the test sample and the negative control sample is an indication of the presence of human PD-L1 antigen in the sample. Various methods can be used to quantify PD-L1 expression.

[0206] In certain embodiments, the automated IHC method includes (a) deparaffinizing and rehydrating mounted tissue sections in an automated stainer; (b) retrieving antigens by heating at 110°C for 10 minutes using a decloaking chamber and pH 6 buffer; (c) placing the reagents in the automated stainer; and (d) running the automated stainer to include the steps of neutralizing endogenous peroxidase in the tissue specimen; blocking nonspecific protein binding sites on the slide; incubating the slide with primary antibody; incubating with primary post-blocking reagent; incubating with NovoLink polymer; adding chromogen substrate and developing; and counterstaining with hematoxylin.

[0207] For the evaluation of PD-L1 expression in tumor tissue samples, pathologists performed membrane PD-L1 analysis within each microscopic field. +The number of tumor cells is tested, and the percentage of positive cells is estimated mentally and then averaged to arrive at the final percentage. Various staining intensities are defined as 0 / negative, 1+ / weak, 2+ / moderate, and 3+ / strong. Generally, percentage values ​​are first assigned to 0 and 3+ buckets, and then intermediate 1+ and 2+ intensities are considered. For highly heterogeneous tissues, the specimen is divided into zones, each zone is scored separately, and then combined into a set of percentage values. The percentages of negative cells and cells positive at various staining intensities are determined from each region, and a median value is obtained for each zone. Final percentage values ​​are obtained for tissue in each staining intensity category: negative, 1+, 2+, and 3+. The sum of all staining intensities should equal 100%. In some embodiments, the threshold number of cells required for PD-L1 positivity is at least about 100, at least about 125, at least about 150, at least about 175, or at least about 200 cells. In one embodiment, the threshold number of cells required to be PD-L1 positive is at least about 100 cells.

[0208] Staining is also assessed in tumor-infiltrating inflammatory cells, such as macrophages and lymphocytes. In most cases, staining is observed in the majority of macrophages, which serve as an internal positive control. While staining at 3+ intensity is not required, the absence of macrophage staining should be considered to rule out any technical failure. Macrophages and lymphocytes are assessed for plasma membrane staining, and all samples are scored as positive or negative for each cell category. Staining is also characterized by an external / internal tumor immune cell designation. "Internal" means that the immune cells are located within the tumor tissue and / or on the tumor area border without any physical intervening tumor cells. "External" means that there is no physical relationship to the tumor, and the immune cells are found in the periphery associated with connective tissue or any related adjacent tissue.

[0209] In some embodiments of these scoring methods, samples are scored by two pathologists working independently, and the scores are then combined. In other embodiments, the identification of positive and negative cells is scored using appropriate software.

[0210] The Histoscore is used as a more quantitative measure of the IHC data and is calculated as follows: Histoscore = [(% tumor × 1 (low intensity)) + (% tumor × 2 (moderate intensity)) + (% tumor × 3 (high intensity)]

[0211] To determine the Histoscore, the pathologist estimates the percentage of staining cells in each intensity category within the specimen. Because expression of most biomarkers is heterogeneous, the Histoscore truly represents overall expression. Final Histoscore ranges from 0 (no expression) to 300 (maximal expression).

[0212] Another means of quantifying PD-L1 expression in test tissue sample IHC is the adjusted inflammation score (AIS), a score defined as the density of inflammation multiplied by the percent PD-L1 expression by tumor-infiltrating inflammatory cells (Taube et al., "Colocalization of inflammatory response with B7-h1 expression in human melanocytic lesions supports an adaptive resistance mechanism of immune escape," Sci. Transl. Med. 4(127):127ra37 (2012)).

[0213] In some embodiments, the PD-L1 expression level of the tumor is at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 11%, at least about 12%, at least about 13%, at least about 14%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100%. In other embodiments, the PD-L1 status of the tumor is at least about 1%. In other embodiments, the PD-L1 status of the subject is at least about 5%. In some embodiments, the PD-L1 status of the tumor is at least about 10%. In some embodiments, the PD-L1 status of the tumor is at least about 25%. In certain embodiments, the PD-L1 status of the tumor is at least about 50%.

[0214] As used herein, "PD-L1 positive" can be used interchangeably with "at least about 1% PD-L1 expression." In certain embodiments, a PD-L1-positive tumor may therefore have at least about 1%, at least about 2%, at least about 5%, at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% of tumor cells expressing PD-L1, as measured by automated IHC. In certain embodiments, "PD-L1 positive" means that there are at least 100 cells that express PD-L1 on the cell surface.

[0215] In certain embodiments, tumors with PD-L1 positive high TMB have a greater likelihood of responding to treatment with an anti-PD-1 antibody than tumors with only high TMB, only PD-L1 positive expression, or neither. In certain embodiments, the tumor has at least about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, or about 50% PD-L1 expression. In certain embodiments, tumors with ≧50% PD-L1 expression and high TMB status are more likely to respond to treatment with an anti-PD-1 antibody than tumors with only high TMB, only ≧50% PD-L1 expression, or neither.

[0216] In certain embodiments, tumors in subjects suitable for immunotherapy, e.g., anti-PD-1 antibody treatment, according to the present invention do not express PD-L1 (less than 1%, less than 2%, less than 3%, less than 4%, or less than 5% membranous PD-L1). In certain embodiments, the methods of the present invention are independent of PD-L1 expression.

[0217] MSI Status TMB status can be used alone or in combination with other factors, such as MSI status, as a means of predicting tumor response to therapy, particularly treatment with cancer immunotherapeutics such as anti-PD-1 antibodies and / or anti-CTLA-4 antibodies. In some embodiments, MSI status is part of TMB status. In other embodiments, MSI status is measured separately from TMB status.

[0218] Microsatellite instability is a genetic hypermutability condition caused by impaired DNA mismatch repair (MMR). The presence of MSI represents phenotypic evidence that MMR is not functioning normally. In most cases, the genetic basis of instability in MSI tumors is inherited germline mutations in any one of five human MMR genes: MSH2, MLH1, MSH6, PMS2, and PMS1. In one embodiment, a subject undergoing tumor (e.g., colon tumor) treatment has high microsatellite instability (MSI-H) and has at least one mutation in the gene MSH2, MLH1, MSH6, PMS2, or PMS1. In another embodiment, a subject undergoing tumor treatment in the control group does not have microsatellite instability (MSS or MSI stable) and does not have a mutation in the gene MSH2, MLH1, MSH6, PMS2, or PMS1.

[0219] In one embodiment, a subject suitable for immunotherapy has a high TMB status and an MSI-H tumor. As used herein, an MSI-H tumor refers to a tumor with at least about 30% or more unstable MSI biomarkers. In one embodiment, the tumor is derived from colorectal cancer. In one embodiment, a tumor is MSI-H colorectal cancer when tumor germline alterations are detected in at least two, at least three, at least four, or at least five MMR genes. In another embodiment, a tumor is MSI-H colorectal cancer when germline alterations are detected in at least 30% of five or more MMR genes. In one embodiment, germline alterations in MMR genes are measured by polymerase chain reaction. In another embodiment, a tumor is MSI-H colorectal cancer when at least one protein encoded by a tumor DNA MMR gene is not detected in the tumor. In one embodiment, at least one protein encoded by a DNA MMR gene is detected by immunohistochemistry.

[0220] Treatment methods of the present invention Certain embodiments of the present invention relate to methods of treating a subject with an SCLC-derived tumor having a high tumor mutational burden (TMB) status, the method comprising administering to the subject an anti-PD-1 antibody or an anti-PD-L1 antibody. Another embodiment of the present invention relates to methods of treating a subject with an SCLC-derived tumor, the method comprising administering to the subject therapeutically effective amounts of an anti-PD-1 antibody (or an anti-PD-L1 antibody) and an antibody or antigen-binding portion thereof that specifically binds to CTLA-4 (an "anti-CTLA-4 antibody"), wherein the tumor has a high TMB status. The method may further comprise measuring the TMB status of a biological sample obtained from the subject. Furthermore, the present invention contemplates administering an anti-PD-1 or anti-PD-L1 antibody to subjects identified as suitable for such treatment, for example, based on measuring a high TMB.

[0221] In certain embodiments, the anti-PD-1 antibody cross-competes with nivolumab for binding to human PD-1. In other embodiments, the anti-PD-1 antibody binds to the same epitope as nivolumab. In certain embodiments, the anti-PD-1 antibody is nivolumab. In other specific embodiments, the anti-PD-1 antibody is pembrolizumab. Additional anti-PD-1 antibodies are described elsewhere herein. In other embodiments, anti-PD-L1 antibodies useful in the methods of the invention are described elsewhere herein.

[0222] In certain embodiments, the anti-PD-1 antibody (or anti-PD-L1 antibody) and / or anti-CTLA-4 antibody is a chimeric antibody, a humanized antibody, a human monoclonal antibody, or an antigen-binding portion thereof. In other embodiments, the anti-PD-1 antibody (or anti-PD-L1 antibody) and anti-CTLA-4 antibody comprise a heavy chain constant region of the human IgG1 or IgG4 isotype.

[0223] In some embodiments, the anti-PD-1 antibody is administered at a dose ranging from 0.1 mg / kg to 10.0 mg / kg body weight every 2, 3, or 4 weeks. In other embodiments, the anti-PD-1 antibody is administered at a dose of 5 mg / kg or 10 mg / kg body weight every 3 weeks. In some embodiments, the anti-PD-1 antibody is administered at a dose of 5 mg / kg body weight every 3 weeks. In other embodiments, the anti-PD-1 antibody is administered at a dose of 3 mg / kg body weight every 2 weeks. In other embodiments, the anti-PD-1 antibody is administered at a dose of 2 mg / kg body weight every 3 weeks.

[0224] In some embodiments, the anti-PD-1 antibody (or anti-PD-L1 antibody) and / or anti-CTLA-4 antibody is administered as a fixed dose. In some embodiments, the anti-PD-1 antibody and / or anti-CTLA-4 antibody is administered as a fixed dose of at least about 200 mg, at least about 220 mg, at least about 240 mg, at least about 260 mg, at least about 280 mg, at least about 300 mg, at least about 320 mg, at least about 340 mg, at least about 360 mg, at least about 380 mg, at least about 400 mg, at least about 420 mg, at least about 440 mg, at least about 460 mg, at least about 480 mg, at least about 500 mg, or at least about 550 mg. In other embodiments, the anti-PD-1 antibody and / or anti-CTLA-4 antibody is administered as a fixed dose about every 1, 2, 3, or 4 weeks.

[0225] In one embodiment, the anti-PD-1 antibody is administered at a dose of 1 mg / kg every three weeks, and the anti-CTLA-4 antibody is administered at a dose of 3 mg / kg every three weeks. In other embodiments, the 1 mg / kg dose of the anti-PD-1 antibody and the 3 mg / kg dose of the anti-CTLA-4 antibody are administered once each, twice each, three times each, four times each, five times each, six times each, seven times each, eight times each, nine times each, or ten times each. In a further embodiment, the combination therapy of the anti-PD-1 antibody and the anti-CTLA-4 antibody is followed by a monotherapy of the anti-PD-1 antibody, e.g., at a dose of 3 mg / kg, every two weeks.

[0226] In certain embodiments, the anti-PD-1 antibody is administered at a subtherapeutic dose. In certain other embodiments, the anti-CTLA-4 antibody is administered at a subtherapeutic dose. In further embodiments, both the anti-PD-1 antibody and the anti-CTLA-4 antibody are each administered at a subtherapeutic dose.

[0227] The present invention provides a method for treating a subject with a tumor derived from SCLC, comprising administering to the subject an anti-PD-1 antibody. The present invention further provides a method for treating a subject with a tumor derived from SCLC, comprising administering to the subject a combination of (a) an anti-PD-1 antibody; and (b) an anti-CTLA-4 antibody. In some embodiments, the subject is a human patient.

[0228] In certain embodiments, the subject is a chemotherapy-naive patient (e.g., a patient who has not previously received any chemotherapy). In other embodiments, the subject has received other cancer treatments (e.g., chemotherapy), but is resistant or refractory to such other cancer treatments. In certain embodiments, the SCLC is recurrent SCLC. In certain embodiments, the subject has received at least one, at least two, at least three, at least four, or at least five prior therapies to treat the tumor. In certain embodiments, the subject has received one prior therapy to treat the tumor. In other embodiments, the subject has received two prior therapies to treat the tumor. In other embodiments, the subject has received three prior therapies to treat the tumor. In other embodiments, the subject has received four prior therapies to treat the tumor. In other embodiments, the subject has received five prior therapies to treat the tumor. In other embodiments, the subject has received more than five prior therapies to treat the tumor.

[0229] In some embodiments, the prior treatment experience includes chemotherapy. In some embodiments, the chemotherapy includes platinum-based therapy. In some embodiments, the platinum-based therapy includes a platinum-based anti-neoplastic agent selected from the group consisting of cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenanthriplatin, picoplatin, satraplatin, and any combination thereof. In certain embodiments, the platinum-based therapy includes cisplatin (e.g., cisplatin in combination with etoposide). In some embodiments, the subject has received prior radiation therapy. In other embodiments, the prior therapy includes antibody therapy.

[0230] In certain specific embodiments, the subject has cancer cells that express a mutant form of the EGFR or KRAS gene. In certain embodiments, the subject has cancer cells that are PD-L1 positive. In certain embodiments, the subject has cancer cells that are PD-L1 negative. In certain embodiments, the subject has never smoked. In certain embodiments, the subject is a former smoker. In certain embodiments, the subject is a current smoker. In certain embodiments, the SCLC comprises small cell carcinoma. In certain embodiments, the SCLC comprises mixed small cell carcinoma.

[0231] Certain types of cancers have a high frequency of mutations and therefore have high TMB (Alexandrov et al., Nature (2013) 500:415-421). Non-limiting examples of cancers with high TMB include melanoma, lung, bladder, and gastrointestinal cancer. In some embodiments, the tumor is lung cancer. In some embodiments, the lung cancer is small cell lung cancer (SCLC). In some embodiments, the NSCLC has squamous histology. In other embodiments, the NSCLC has non-squamous histology. In other embodiments, the tumor is selected from renal cell carcinoma, ovarian cancer, colorectal cancer, gastrointestinal cancer, esophageal cancer, bladder cancer, lung cancer, and melanoma. It should be understood that the methods disclosed herein encompass solid tumors as well as hematological cancers.

[0232] The treatment methods disclosed herein may improve clinical response and / or clinical benefit in subjects with tumors, particularly those with tumors with high TMB. High TMB may correlate with neoantigen load, i.e., neoantigen number and T cell reactivity, and thus immune-mediated anti-tumor response. Thus, high TMB is a factor that can be used alone or in combination with other factors to identify tumors (and patients with such tumors) that are likely to benefit from treatment with anti-PD-1 antibodies and / or anti-PD-L1 antibodies, for example, compared to the current standard of care.

[0233] In certain embodiments, the subject exhibits progression-free survival of at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 1 year, at least about 18 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years after administration. In other embodiments, the subject exhibits overall survival of at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 1 year, at least about 18 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years after administration. In still other embodiments, subjects exhibit an objective response rate of at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100%.

[0234] Anti-PD-1 treatment One embodiment of the present invention provides a method of treating a subject having an SCLC-derived tumor with a high tumor mutational burden (TMB) status, comprising administering to the subject an immunotherapeutic agent comprising an anti-PD-1 antibody (or an anti-PD-L1 antibody) alone, or an anti-PD-1 antibody and an anti-CTLA-4 antibody.

[0235] In certain embodiments, the anti-PD-1 antibody cross-competes with nivolumab for binding to human PD-1. In other embodiments, the anti-PD-1 antibody binds to the same epitope as nivolumab. In other embodiments, anti-PD-1 antibodies useful in the methods of the invention are described elsewhere herein. In other embodiments, an anti-PD-L1 antibody can be used instead of the anti-PD-1 antibody. Non-limiting examples of anti-PD-L1 antibodies are disclosed elsewhere herein.

[0236] In certain embodiments, the anti-PD-1 or anti-PD-L1 antibody is a chimeric antibody, a humanized antibody, a human antibody, or an antigen-binding portion thereof. In other embodiments, the anti-PD-1 or anti-PD-L1 antibody comprises a heavy chain constant region of the human IgG1 or human IgG4 isotype.

[0237] Anti-PD-1 antibodies useful in the present invention Anti-PD-1 antibodies known in the art can be used in the compositions and methods described herein. Various human monoclonal antibodies that specifically bind to PD-1 with high affinity are disclosed in U.S. Pat. No. 8,008,449. The anti-PD-1 human antibodies disclosed in U.S. Pat. No. 8,008,449 have been shown to exhibit one or more of the following properties: (a) a specific binding affinity of 1×10 to human PD-1 as determined by surface plasmon resonance using a Biacore biosensor system; -7 K below M D(b) does not substantially bind to human CD28, CTLA-4, or ICOS; (c) increases T-cell proliferation in a mixed lymphocyte reaction (MLR) assay; (d) increases interferon-γ production in an MLR assay; (e) increases IL-2 secretion in an MLR assay; (f) binds to human PD-1 and cynomolgus PD-1; (g) inhibits the binding of PD-L1 and / or PD-L2 to PD-1; (h) stimulates an antigen-specific memory response; (i) stimulates an antibody response; and (j) inhibits tumor cell growth in vivo. Anti-PD-1 antibodies useful in the present invention are monoclonal antibodies that specifically bind to human PD-1 and exhibit at least one, and in certain embodiments, at least five, of the foregoing properties.

[0238] Other anti-PD-1 monoclonal antibodies are described in, e.g., U.S. Patents 6,808,710, 7,488,802, 8,168,757, and 8,354,509, U.S. Publication No. 2016 / 0272708, and PCT Publication Nos. WO2012 / 145493, WO2008 / 156712, WO2015 / 112900, WO2012 / 145493, WO2015 / 112800, WO2014 / 206107, WO2015 / 35606, WO2015 / 085847, WO2014 / 179664, WO2017 / 020291, WO2017 / 020858, WO2016 / 19736 7, WO2017 / 024515, WO2017 / 025051, WO2017 / 123557, WO2016 / 106159, WO2014 / 194302, WO2017 / 040790, WO2017 / 133540, WO2017 / 132827, WO2017 / 024465, WO2017 / 025016, WO2017 / 106061, WO2017 / 19846, WO2017 / 024465, WO2017 / 025016, WO2017 / 132825 and WO2017 / 133540, each of which is incorporated herein by reference in its entirety.

[0239] In certain embodiments, the anti-PD-1 antibody is selected from the group consisting of nivolumab (OPDIVO®, 5C4, BMS-936558, MDX-1106, and ONO-4538), pembrolizumab (Merck; also known as KEYTRUDA®, lambrolizumab, and MK-3475; see WO2008 / 156712), PDR001 (Novartis; also known as spartalizumab; see WO2015 / 112900), MEDI-0680 (AstraZeneca; also known as AMP-514; see WO2012 / 145493), cemiplimab (Regeneron; also known as REGN-2810; see WO2015 / 112800), JS001 (TAIZHOU JUNSHI PHARMA; Si-Yang Liu et al., J. Hematol. Oncol. 10:136 (2017)), BGB-A317 (tislelizumab; Beigene; see WO2015 / 35606 and US2015 / 0079109), INCSHR1210 (Jiangsu Hengrui Medicine; also known as SHR-1210; see WO2015 / 085847; Si-Yang Liu et al., J. Hematol. Oncol. 10:136 (2017)), TSR-042 (Tesaro Biopharmaceutical; also known as ANB011; see WO2014 / 179664), GLS-010 (Wuxi / Harbin Gloria Pharmaceuticals; also known as WBP3055; Si-Yang Liu et al., J. Hematol. Oncol. 10:136 (2017)), AM-0001 (Armo), STI-1110 (Sorrento Therapeutics; see WO2014 / 194302), AGEN2034 (Agenus; see WO2017 / 040790), MGA012 (Macrogenics; see WO2017 / 19846), IBI308 (Innovent; see WO2017 / 024465, WO2017 / 025016, WO2017 / 132825 and WO2017 / 133540), and BCD-100 (Biocad).

[0240] In one embodiment, the anti-PD-1 antibody is nivolumab, a fully human IgG4(S228P) PD-1 immune checkpoint inhibitor antibody that selectively blocks interaction with PD-1 ligands (PD-L1 and PD-L2), thereby blocking downregulation of anti-tumor T cell function (US Pat. No. 8,008,449; Wang et al., 2014 Cancer Immunol Res. 2(9):846-56).

[0241] In another embodiment, the anti-PD-1 antibody is pembrolizumab. Pembrolizumab is a humanized monoclonal IgG4 (S228P) antibody directed against the human cell surface receptor PD-1 (programmed cell death-1 or planned cell death-1). Pembrolizumab is described, for example, in U.S. Patents 8,354,509 and 8,900,587.

[0242] Anti-PD-1 antibodies useful in the disclosed compositions and methods also include isolated antibodies that specifically bind to human PD-1 and cross-compete with any of the anti-PD-1 antibodies disclosed herein, e.g., nivolumab, for binding to human PD-1 (see, e.g., U.S. Pat. Nos. 8,008,449 and 8,779,105; WO 2013 / 173223). In certain embodiments, the anti-PD-1 antibody binds to the same epitope as any of the anti-PD-1 antibodies described herein, e.g., nivolumab. The ability of antibodies to cross-compete for binding to an antigen indicates that these monoclonal antibodies bind to the same epitope region of the antigen and sterically hinder the binding of other cross-competing antibodies to that particular epitope region. These cross-competing antibodies are expected to have functional properties very similar to those of a control antibody, e.g., nivolumab, due to their binding to the same epitope region of PD-1. Cross-competing antibodies can be readily identified based on their ability to cross-compete with nivolumab in standard PD-1 binding assays, such as Biacore analysis, ELISA assays, or flow cytometry (see, e.g., WO2013 / 173223).

[0243] In some embodiments, the human PD-1 antibody, the antibody that cross-competes with nivolumab for binding to human PD-1 or binds to the same epitope region as nivolumab, is a monoclonal antibody. For administration to human subjects, these cross-competing antibodies are chimeric, engineered, or humanized or human antibodies. Such chimeric, engineered, humanized, or human monoclonal antibodies can be prepared and isolated by methods well known in the art.

[0244] Anti-PD-1 antibodies useful in the compositions and methods of the invention also include antigen-binding portions of such antibodies. It is well established that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody.

[0245] Anti-PD-1 antibodies suitable for use in the disclosed compositions and methods are those that bind to PD-1 with high specificity and affinity, block binding of PD-L1 and / or PD-L2, and inhibit the immunosuppressive effects of the PD-1 signaling pathway. In any of the compositions or methods disclosed herein, an anti-PD-1 "antibody" includes an antigen-binding portion or fragment that binds to the PD-1 receptor and exhibits functional properties similar to those of whole antibodies with respect to ligand binding inhibition and immune system upregulation. In certain embodiments, the anti-PD-1 antibody or antigen-binding portion thereof cross-competes with nivolumab for binding to human PD-1.

[0246] In some embodiments, the anti-PD-1 antibody is administered at a dose ranging from 0.1 mg / kg to 20.0 mg / kg body weight every 2, 3, 4, 5, 6, 7, or 8 weeks, e.g., 0.1 mg / kg to 10.0 mg / kg body weight every 2, 3, or 4 weeks. In other embodiments, the anti-PD-1 antibody is administered at a dose of about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, or 10 mg / kg body weight every 2 weeks. In other embodiments, the anti-PD-1 antibody is administered at a dose of about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, or 10 mg / kg body weight every 3 weeks. In one embodiment, the anti-PD-1 antibody is administered at a dose of about 5 mg / kg body weight about every three weeks. In another embodiment, the anti-PD-1 antibody, e.g., nivolumab, is administered at a dose of about 3 mg / kg body weight about every two weeks. In another embodiment, the anti-PD-1 antibody, e.g., pembrolizumab, is administered at a dose of about 2 mg / kg body weight about every three weeks.

[0247] The anti-PD-1 antibodies useful in the present invention may be administered in a fixed dose. In certain embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 100 to about 1000 mg, about 100 mg to about 900 mg, about 100 mg to about 800 mg, about 100 mg to about 700 mg, about 100 mg to about 600 mg, about 100 mg to about 500 mg, about 200 mg to about 1000 mg, about 200 mg to about 900 mg, about 200 mg to about 800 mg, about 200 mg to about 700 mg, about 200 mg to about 600 mg, about 200 mg to about 500 mg, about 200 mg to about 480 mg, or about 240 mg to about 480 mg; in certain embodiments, the anti-PD-1 antibody is administered at an administration interval of about 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, or 10 weeks. at least about 260 mg, at least about 280 mg, at least about 300 mg, at least about 320 mg, at least about 340 mg, at least about 360 mg, at least about 380 mg, at least about 400 mg, at least about 420 mg, at least about 440 mg, at least about 460 mg, at least about 480 mg, at least about 500 mg, at least about 520 mg, at least about 540 mg, at least about 550 mg, at least about 560 mg, at least about 580 mg, at least about 600 mg, at least about 620 mg, at least about 640 mg, at least about 660 mg, at least about 680 mg, at least about 700 mg or at least about 720 mg. In other embodiments, the anti-PD-1 antibody is administered at a fixed dose of about 200 mg to about 800 mg, about 200 mg to about 700 mg, about 200 mg to about 600 mg, or about 200 mg to about 500 mg at an administration interval of about 1 week, 2 weeks, 3 weeks, or 4 weeks.

[0248] In one embodiment, the anti-PD-1 antibody is administered at a flat dose of about 200 mg about once every three weeks. In another embodiment, the anti-PD-1 antibody is administered at a flat dose of about 200 mg about once every two weeks. In another embodiment, the anti-PD-1 antibody is administered at a flat dose of about 240 mg about once every two weeks. In one embodiment, the anti-PD-1 antibody is administered at a flat dose of about 480 mg about once every four weeks.

[0249] Anti-PD-L1 antibodies useful in the present invention Because anti-PD-1 and anti-PD-L1 target the same signaling pathway and have been shown in clinical trials to exhibit similar levels of efficacy in a variety of cancers, including renal cell carcinoma (see Brahmer et al. (2012) N Engl J Med 366:2455-65; Topalian et al. (2012a) N Engl J Med 366:2443-54; WO2013 / 173223), anti-PD-L1 antibodies can be substituted for anti-PD-1 antibodies in any of the therapeutic methods disclosed herein. Accordingly, certain aspects of the invention include: The present invention relates to a method of treating a subject with a tumor with a high TMB status, e.g., SCLC, comprising administering to the subject an anti-PD-L1 antibody alone ("monotherapy") or an anti-PD-L1 antibody in combination with an anti-CTLA-4 antibody. Anti-PD-L1 antibodies known in the art can be used in the compositions and methods of the invention. Examples of anti-PD-L1 antibodies useful in the compositions and methods of the invention include the antibodies disclosed in U.S. Patent 9,580,507. The anti-PD-L1 human monoclonal antibody disclosed in U.S. Patent 9,580,507 has been shown to exhibit one or more of the following properties: (a) a specific affinity for human PD-L1 of 1x10 as determined by surface plasmon resonance using a Biacore biosensor system; -7 K below M D (b) increase T cell proliferation in a mixed lymphocyte reaction (MLR) assay; (c) increase interferon-γ production in an MLR assay; (d) increase IL-2 secretion in an MLR assay; (e) stimulate antibody responses; and (f) reverse the effects of T regulatory cells on T cell effector cells and / or dendritic cells. Anti-PD-L1 antibodies useful in the present invention include monoclonal antibodies that specifically bind to human PD-L1 and exhibit at least one, and in certain embodiments, at least five, of the foregoing properties.

[0250] In some embodiments, the anti-PD-L1 antibody is selected from the group consisting of BMS-936559 (12A4, also known as MDX-1105; see, e.g., US 7,943,743 and WO 2013 / 173223), atezolizumab (Roche; Tecentriq®; also known as MPDL3280A, RG7446; see US 8,217,149; see also Herbst et al. (2013) J Clin Oncol 31(suppl):3000), durvalumab (AstraZeneca; Imfinzi) TM , also known as MEDI-4736; see WO2011 / 066389), avelumab (Pfizer; also known as BAVENCIO®, MSB-0010718C; see WO2013 / 079174), STI-1014 (Sorrento; see WO2013 / 181634), CX-072 (Cytomx; see WO2016 / 149201), KN035 (3D Med / Alphamab; see Zhang et al., Cell Discov. 7:3 (March 2017)), LY3300054 (Eli Lilly Co.; see, e.g., WO2017 / 034916), and CK-301 (Checkpoint Therapeutics; Gorelik et al., AACR:Abstract 4606 (Apr 2016).

[0251] In one embodiment, the PD-L1 antibody is atezolizumab (Tecentriq®). Atezolizumab is a fully humanized IgG1 monoclonal anti-PD-L1 antibody.

[0252] In one embodiment, the PD-L1 antibody is durvalumab (Imfinzi). TM Durvalumab is a human IgG1 kappa monoclonal anti-PD-L1 antibody.

[0253] In one embodiment, the PD-L1 antibody is avelumab (BAVENCIO®). Avelumab is a human IgG1 lambda monoclonal anti-PD-L1 antibody.

[0254] Anti-PD-L1 antibodies useful in the disclosed compositions and methods also include isolated antibodies that specifically bind to human PD-L1 and cross-compete with any of the anti-PD-L1 antibodies disclosed herein, e.g., atezolizumab, durvalumab, and / or avelumab, for binding to human PD-L1. In certain embodiments, the anti-PD-L1 antibody binds to the same epitope as any of the anti-PD-L1 antibodies described herein, e.g., atezolizumab, durvalumab, and / or avelumab. The ability of antibodies to cross-compete for binding to an antigen indicates that these antibodies bind to the same epitope region of the antigen and sterically interfere with the binding of other cross-competing antibodies to that particular epitope region. These cross-competing antibodies are expected to have functional properties very similar to those of control antibodies, e.g., atezolizumab and / or avelumab, due to their binding to the same epitope region of PD-L1. Cross-competing antibodies can be readily identified based on their ability to cross-compete with atezolizumab and / or avelumab in standard PD-L1 binding assays, such as Biacore analysis, ELISA assays or flow cytometry (see, e.g., WO2013 / 173223).

[0255] In some embodiments, antibodies that cross-compete with human PD-L1 antibodies, such as atezolizumab, durvalumab, and / or avelumab, for binding to human PD-L1 or that bind to the same epitope region are monoclonal antibodies. For administration to human subjects, these cross-competing antibodies are chimeric, engineered, or humanized or human antibodies. Such chimeric, engineered, humanized, or human monoclonal antibodies can be prepared and isolated by methods well known in the art.

[0256] Anti-PD-L1 antibodies useful in the compositions and methods of the invention also include antigen-binding portions of such antibodies. It is well established that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody.

[0257] Anti-PD-L1 antibodies suitable for use in the disclosed compositions and methods are those that bind to PD-L1 with high specificity and affinity, block PD-1 binding, and inhibit the immunosuppressive effects of the PD-1 signaling pathway. In any of the compositions or methods disclosed herein, an anti-PD-L1 "antibody" includes an antigen-binding portion or fragment that binds to PD-L1 and exhibits functional properties similar to those of whole antibodies in receptor binding inhibition and immune system upregulation. In certain embodiments, the anti-PD-L1 antibody or antigen-binding portion thereof cross-competes with atezolizumab, durvalumab, and / or avelumab for binding to human PD-L1.

[0258] The anti-PD-L1 antibody useful in the present invention can be any PD-L1 antibody that specifically binds to PD-L1, for example, an antibody that cross-competes with durvalumab, avelumab, or atezolizumab for binding to human PD-1, for example, an antibody that binds to the same epitope as durvalumab, avelumab, or atezolizumab. In certain embodiments, the anti-PD-L1 antibody is durvalumab. In other embodiments, the anti-PD-L1 antibody is avelumab. In some embodiments, the anti-PD-L1 antibody is atezolizumab.

[0259] In some embodiments, the anti-PD-L1 antibody is administered at a dose ranging from about 0.1 mg / kg to about 20.0 mg / kg body weight, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 11 mg / kg, about 12 mg / kg, about 13 mg / kg, about 14 mg / kg, about 15 mg / kg, about 16 mg / kg, about 17 mg / kg, about 18 mg / kg, about 19 mg / kg, or about 20 mg / kg, about every 2, 3, 4, 5, 6, 7, or 8 weeks.

[0260] In one embodiment, the anti-PD-L1 antibody is administered at a dose of about 15 mg / kg body weight about every three weeks. In another embodiment, the anti-PD-L1 antibody is administered at a dose of about 10 mg / kg body weight about every two weeks.

[0261] In other embodiments, the anti-PD-L1 antibodies useful in the present invention are administered at a fixed dose of about 200 mg to about 1600 mg, about 200 mg to about 1500 mg, about 200 mg to about 1400 mg, about 200 mg to about 1300 mg, about 200 mg to about 1200 mg, about 200 mg to about 1100 mg, about 200 mg to about 1000 mg, about 200 mg to about 900 mg, about 200 mg to about 800 mg, about 200 mg to about 700 mg, about 200 mg to about 600 mg, about 700 mg to about 1300 mg, about 800 mg to about 1200 mg, about 700 mg to about 900 mg, or about 1100 mg to about 1300 mg. In certain embodiments, the anti-PD-L1 antibody is administered at a fixed dose of at least about 240 mg, at least about 300 mg, at least about 320 mg, at least about 400 mg, at least about 480 mg, at least about 500 mg, at least about 560 mg, at least about 600 mg, at least about 640 mg, at least about 700 mg, at least about 720 mg, at least about 800 mg, at least about 880 mg, at least about 900 mg, at least 960 mg, at least about 1000 mg, at least about 1040 mg, at least about 1100 mg, at least about 1120 mg, at least about 1200 mg, at least about 1280 mg, at least about 1300 mg, at least about 1360 mg, or at least about 1400 mg, with a dosing interval of about 1 week, 2 weeks, 3 weeks, or 4 weeks. In certain embodiments, the anti-PD-L1 antibody is administered at a fixed dose of about 1000 mg. In some embodiments, the anti-PD-L1 antibody is administered at a fixed dose of about 1100 mg. In some embodiments, the anti-PD-L1 antibody is administered at a fixed dose of about 1200 mg. In some embodiments, the anti-PD-L1 antibody is administered at a fixed dose of about 1300 mg. In some embodiments, the anti-PD-L1 antibody is administered at a fixed dose of about 1400 mg. In some embodiments, the anti-PD-L1 antibody is administered at a fixed dose of about 1500 mg. In some embodiments, the anti-PD-L1 antibody is administered at a fixed dose of about 1200 mg about every three weeks. In other embodiments, the anti-PD-L1 antibody is administered at a fixed dose of about 800 mg about every two weeks.

[0262] Anti-CTLA-4 antibody Anti-CTLA-4 antibodies known in the art can be used in the compositions and methods of the present invention. The anti-CTLA-4 antibodies of the present invention bind to human CTLA-4 in a manner that disrupts the interaction between CTLA-4 and human B7 receptors. Because the interaction between CTLA-4 and B7 transmits a signal that inactivates T cells bearing the CTLA-4 receptor, disrupting the interaction effectively induces, enhances, or prolongs the activation of such T cells, thereby inducing, enhancing, or prolonging an immune response.

[0263] Human monoclonal antibodies that specifically bind to CTLA-4 with high affinity are disclosed in U.S. Patent 6,984,720. Other anti-CTLA-4 monoclonal antibodies are described, for example, in U.S. Patents 5,977,318, 6,051,227, 6,682,736, and 7,034,121 and International Publication Nos. WO 2012 / 122444, WO 2007 / 113648, WO 2016 / 196237, and WO 2000 / 037504, each of which is incorporated herein by reference in its entirety. The anti-CTLA-4 human monoclonal antibodies disclosed in U.S. Patent 6,984,720 have been shown to exhibit one or more of the following properties: (a) a specific binding affinity to human CTLA-4 of at least about 10, as determined by Biacore analysis; 7 M -1 or about 10 9 M -1 or about 10 10 M -1 ~10 11 M -1 Equilibrium binding constant (K a (b) specifically binds with a binding affinity reflected by at least about 10 3 , about 10 4 or about 10 5 m -1 s -1 The binding rate constant (k a );(c) at least about 10 3 , about 10 4 or about 10 5 m -1 s -1 The bond dissociation constant (k dand (d) inhibit the binding of CTLA-4 to B7-1 (CD80) and B7-2 (CD86). Anti-CTLA-4 antibodies useful in the present invention are monoclonal antibodies that specifically bind to human CTLA-4 and exhibit at least one, at least two, or at least three of the above properties.

[0264] In certain embodiments, the CTLA-4 antibody is selected from the group consisting of ipilimumab (Yervoy®, also known as MDX-010, 10D1; see U.S. Pat. No. 6,984,720), MK-1308 (Merck), AGEN-1884 (Agenus Inc.; see WO2016 / 196237), and tremelimumab (AstraZeneca; also known as ticilimumab, CP-675,206; WO2000 / 037504 and Ribas, Update Cancer Ther. 2(3): 133-39 (2007)). In certain embodiments, the anti-CTLA-4 antibody is ipilimumab.

[0265] In certain embodiments, the CTLA-4 antibody is ipilimumab for use in the compositions and methods disclosed herein. Ipilimumab is a fully human, IgG1 monoclonal antibody that blocks the binding of CTLA-4 to its B7 ligand, thereby stimulating T cell activation and improving overall survival (OS) in patients with advanced melanoma.

[0266] In certain embodiments, the CTLA-4 antibody is tremelimumab.

[0267] In certain embodiments, the CTLA-4 antibody is MK-1308.

[0268] In certain embodiments, the CTLA-4 antibody is AGEN-1884.

[0269] Anti-CTLA-4 antibodies useful in the disclosed compositions and methods also include isolated antibodies that specifically bind to human CTLA-4 and cross-compete with any of the anti-CTLA-4 antibodies disclosed herein, e.g., ipilimumab and / or tremelimumab, for binding to human CTLA-4. In certain embodiments, the anti-CTLA-4 antibody binds to the same epitope as any of the anti-CTLA-4 antibodies described herein, e.g., ipilimumab and / or tremelimumab. The ability of antibodies to cross-compete for binding to an antigen indicates that these antibodies bind to the same epitope region of the antigen and sterically block the binding of other cross-competing antibodies to that particular epitope region. These cross-competing antibodies are expected to have functional properties very similar to those of control antibodies, e.g., ipilimumab and / or tremelimumab, due to their binding to the same epitope region of CTLA-4. Cross-competing antibodies can be readily identified based on their ability to cross-compete with ipilimumab and / or tremelimumab in standard CTLA-4 binding assays such as Biacore analysis, ELISA assays or flow cytometry (see, for example, WO2013 / 173223).

[0270] In some embodiments, the antibody that cross-competes with human CTLA-4 antibodies such as ipilimumab and / or tremelimumab for binding to human CTLA-4 or binds to the same epitope region is a monoclonal antibody.For administration to human subjects, these cross-competing antibodies are chimeric antibodies, modified antibodies, or humanized or human antibodies.Such chimeric, modified, humanized or human monoclonal antibodies can be prepared and isolated by methods well known in the art.

[0271] Anti-CTLA-4 antibodies useful in the compositions and methods of the invention also include antigen-binding portions of such antibodies. It is well established that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody.

[0272] Anti-CTLA-4 antibodies suitable for use in the disclosed methods or compositions are those that bind to CTLA-4 with high specificity and affinity, block the activity of CTLA-4, and interfere with the interaction of CTLA-4 with human B7 receptors. In any of the compositions or methods disclosed herein, the anti-CTLA-4 "antibody" includes an antigen-binding portion or fragment that binds to CTLA-4 and exhibits functional properties similar to those of the whole antibody with respect to inhibiting the interaction of CTLA-4 with human B7 receptors and upregulating the immune system. In one embodiment, the anti-CTLA-4 antibody or antigen-binding portion thereof cross-competes with ipilimumab and / or tremelimumab for binding to human CTLA-4.

[0273] In some embodiments, the anti-CTLA-4 antibody or antigen-binding portion thereof is administered at a dose ranging from 0.1 mg / kg to 10.0 mg / kg body weight every 2, 3, 4, 5, 6, 7, or 8 weeks. In some embodiments, the anti-CTLA-4 antibody or antigen-binding portion thereof is administered at a dose of 1 mg / kg or 3 mg / kg body weight every 3, 4, 5, or 6 weeks. In some embodiments, the anti-CTLA-4 antibody or antigen-binding portion thereof is administered at a dose of 3 mg / kg body weight every 2 weeks. In other embodiments, the anti-PD-1 antibody or antigen-binding portion thereof is administered at a dose of 1 mg / kg body weight every 6 weeks.

[0274] In certain embodiments, the anti-CTLA-4 antibody, or antigen-binding portion thereof, is administered as a fixed dose of about 10 to about 1,000 mg, about 10 to about 900 mg, about 10 to about 800 mg, about 10 to about 700 mg, about 10 to about 600 mg, about 10 to about 500 mg, about 100 mg to about 1,000 mg, about 100 mg to about 900 mg, about 100 mg to about 800 mg, about 100 mg to about 700 mg, about 100 mg to about 100 mg, about 100 mg to about 500 mg, about 100 mg to about 480 mg, or about 240 mg to about 480 mg. In certain embodiments, the anti-CTLA-4 antibody or antigen-binding portion thereof is at least about 60 mg, at least about 80 mg, at least about 100 mg, at least about 120 mg, at least about 140 mg, at least about 160 mg, at least about 180 mg, at least about 200 mg, at least about 220 mg, at least about 240 mg, at least about 260 mg, at least about 280 mg, at least about 300 mg, at least about 320 mg, at least about 340 mg, at least about 360 mg, at least about 380 mg, at least about 400 mg, at least about 420 mg, at least about 440 mg, at least about 460 mg, at least about 480 mg, at least about 490 mg, at least about 500 mg, at least about 510 mg, at least about 520 mg, at least about 530 mg, at least about 540 mg, at least about 550 mg, at least about 560 mg, at least about 570 mg, at least about 580 mg, at least about 590 mg, at least about 600 mg, at least about 610 mg, at least about 620 mg, at least about 630 mg, at least about 640 mg, at least about 650 mg, at least about 660 mg, at least about 670 mg, at least about 680 mg, at least about 690 mg, at least about 700 mg, at least about 710 mg, at least about 720 mg, at least about 730 mg, at least about 740 mg, at least about 750 mg, at least about 760 mg, at least about 770 mg, at least about 780 mg, at least about 790 mg, at least about 800 mg, at least about 810 mg In another embodiment, the anti-CTLA-4 antibody or antigen-binding portion thereof is administered as a fixed dose of at least about 400 mg, at least about 420 mg, at least about 440 mg, at least about 460 mg, at least about 480 mg, at least about 500 mg, at least about 520 mg, at least about 540 mg, at least about 550 mg, at least about 560 mg, at least about 580 mg, at least about 600 mg, at least about 620 mg, at least about 640 mg, at least about 660 mg, at least about 680 mg, at least about 700 mg, or at least about 720 mg. In another embodiment, the anti-CTLA-4 antibody or antigen-binding portion thereof is administered as a fixed dose about every 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, or 8 weeks.

[0275] cytokines In some embodiments, the method comprises administering a combination therapy comprising (a) an anti-PD-1 antibody and a cytokine or (b) an anti-PD-1 antibody, an anti-CTLA-4 antibody, and a cytokine. The cytokine can be any cytokine known in the art or a variant thereof. In some embodiments, the cytokine is selected from the group consisting of interleukin-2 (IL-2), IL-1β, IL-6, TNF-α, RANTES, monocyte chemoattractant protein (MCP-1), monocyte inflammatory proteins (MIP-1α and MIP-1β), IL-8, lymphotactin, fractalkine, IL-1, IL-4, IL-10, IL-11, IL-13, LIF, interferon-alpha, TGF-beta, and any combination thereof. In some embodiments, the cytokine is a CD122 agonist. In some embodiments, the cytokine comprises IL-2 or a variant thereof.

[0276] In certain embodiments, the cytokine comprises one or more amino acid substitutions, deletions, or insertions compared to the wild-type cytokine amino acid sequence, hi certain embodiments, the cytokine comprises an amino acid sequence in which at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 amino acids are substituted compared to the amino acid sequence of the wild-type cytokine.

[0277] In some embodiments, cytokines are modified, for example, to increase activity and / or half-life. In some embodiments, cytokines are modified by fusing a heterologous moiety to the cytokine. The heterologous moiety can be any structure, including a polypeptide, a polymer, a small molecule, a nucleotide, or a fragment or analog thereof. In some embodiments, the heterologous moiety comprises a polypeptide. In some embodiments, the heterologous moiety comprises albumin or a fragment thereof, albumin-binding polypeptide (ABP), XTEN, Fc, PAS, the C-terminal peptide (CTP) of the beta subunit of human chorionic gonadotropin, or any combination thereof.

[0278] In some embodiments, the cytokine is modified by fusing a polymer to the cytokine. In some embodiments, the polymer comprises polyethylene glycol (PEG), polypropylene glycol (PPG), hydroxyethyl starch (HES), or any combination thereof. As used herein, "PEG" or "polyethylene glycol" includes any water-soluble poly(ethylene oxide). Unless otherwise specified, a "PEG polymer" or polyethylene glycol is one in which substantially all (preferably all) of the monomeric subunits are ethylene oxide subunits, although the polymer may contain different end-capping moieties or functional groups, e.g., for conjugation. PEG polymers for use in the present invention may have the following two structures: "-(CH2CHO) n-n or "-(CH2CH2O) n-1 As noted above, for PEG polymers, the variable (n) ranges from about 3 to 4000, and the end groups and overall structure of the PEG can vary.

[0279] In certain embodiments, the methods of the present invention comprise administering to a subject with a high TMB status (a) an anti-PD-1 antibody and a CD122 agonist, or (b) an anti-PD-1 antibody, an anti-CTLA-4 antibody, and a CD122 agonist. In certain embodiments, the immunotherapy comprises administering (1) an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-CTLA-4 antibody, or any combination thereof, and (2) a CD122 agonist. In certain embodiments, the CD122 agonist comprises IL-2 or a variant thereof. In certain embodiments, the CD122 agonist comprises an IL-2 variant having at least one amino acid substitution compared to wild-type IL-2. In certain embodiments, the CD122 agonist comprises IL-2 fused to PEG. In certain embodiments, the CD122 agonist comprises an IL-2 variant having at least one amino acid substitution compared to wild-type IL-2, wherein the IL-2 variant is fused to PEG.

[0280] Other immunotherapies In some embodiments of the present invention, the methods disclosed herein further comprise administering an additional anti-cancer therapy. In some embodiments, the additional anti-cancer therapy comprises immunotherapy. In some embodiments, the additional anti-cancer therapy comprises administering an antibody or antigen-binding portion thereof that specifically binds to LAG3, TIGIT, TIM3, NKG2a, OX40, ICOS, MICA, CD137, KIR, TGFβ, IL-10, IL-8, B7-H4, Fas ligand, CXCR4, mesothelin, CD27, GITR, or any combination thereof.

[0281] Anti-LAG-3 antibody One embodiment of the present invention relates to a method of treating a subject having a tumor with high TMB status, comprising administering to the subject an immunotherapeutic agent, wherein the immunotherapeutic agent comprises an anti-LAG-3 antibody or antigen-binding portion thereof. The method can further include measuring the TMB status of a biological sample from the subject. Furthermore, the present invention contemplates administering the anti-LAG-3 antibody or antigen-binding portion thereof to a subject identified as suitable for such treatment, e.g., based on measuring high TMB.

[0282] The anti-LAG-3 antibody of the present disclosure binds to human LAG-3. Antibodies that bind to LAG-3 are disclosed in International Publication WO / 2015 / 042246 and U.S. Publications 2014 / 0093511 and 2011 / 0150892. An example of a LAG-3 antibody useful in the present invention is 25F7 (described in U.S. Publication 2011 / 0150892). A further exemplary LAG-3 antibody useful in the present invention is BMS-986016. In certain embodiments, the anti-LAG-3 antibody useful in the composition cross-competes with 25F7 or BMS-986016. In other embodiments, the anti-LAG-3 antibody useful in the composition binds to the same epitope as 25F7 or BMS-986016. In other embodiments, the anti-LAG-3 antibody comprises the six CDRs of 25F7 or BMS-986016.

[0283] Anti-CD137 antibody One embodiment of the present invention relates to a method of treating a subject having a tumor with high TMB status, comprising administering to the subject an immunotherapeutic agent, wherein the immunotherapeutic agent comprises an anti-CD137 antibody or antigen-binding portion thereof. The method can further include measuring the TMB status of a biological sample from the subject. Additionally, the present invention contemplates administering the anti-CD137 antibody or antigen-binding portion thereof to a subject identified as suitable for such treatment, e.g., based on measuring high TMB.

[0284] Anti-CD137 antibodies specifically bind to and activate CD137-expressing immune cells, stimulating immune responses, particularly cytotoxic T cell responses, against tumor cells. Antibodies that bind to CD137 are disclosed in U.S. Publication No. 2005 / 0095244 and U.S. Patents 7,288,638, 6,887,673, 7,214,493, 6,303,121, 6,569,997, 6,905,685, 6,355,476, 6,362,325, 6,974,863, and 6,210,669.

[0285] In some embodiments, the anti-CD137 antibody is urelumab (BMS-663513), described in U.S. Patent 7,288,638 (20H4.9-IgG4 [10C7 or BMS-663513]). In some embodiments, the anti-CD137 antibody is BMS-663031 (20H4.9-IgG1), described in U.S. Patent 7,288,638. In some embodiments, the anti-CD137 antibody is 4E9 or BMS-554271, described in U.S. Patent 6,887,673. In some embodiments, the anti-CD137 antibody is an antibody disclosed in U.S. Patent 7,214,493; 6,303,121; 6,569,997; 6,905,685; or 6,355,476. In some embodiments, the anti-CD137 antibody is 1D8 or BMS-469492; 3H3 or BMS-469497; or 3E1 described in U.S. Patent 6,362,325. In some embodiments, the anti-CD137 antibody is an antibody disclosed in U.S. Patent 6,974,863 (e.g., 53A2). In some embodiments, the anti-CD137 antibody is an antibody disclosed in U.S. Patent 6,210,669 (e.g., 1D8, 3B8, or 3E1). In some embodiments, the antibody is Pfizer's PF-05082566 (PF-2566). In other embodiments, the anti-CD137 antibody useful in the present invention cross-competes with the anti-CD137 antibodies disclosed herein. In some embodiments, the anti-CD137 antibody binds to the same epitope as the anti-CD137 antibodies disclosed herein. In other embodiments, an anti-CD137 antibody useful in the invention comprises six CDRs of an anti-CD137 antibody disclosed herein.

[0286] Anti-KIR antibody

[0010] Certain aspects of the present invention relate to methods of treating a subject with a tumor having high TMB status, comprising administering to the subject an immunotherapeutic agent, wherein the immunotherapeutic agent comprises an anti-KIR antibody or antigen-binding portion thereof. The method may further comprise measuring the TMB status of a biological sample obtained from the subject. Furthermore, the present invention contemplates administering the anti-KIR antibody or antigen-binding portion thereof to a subject identified as suitable for such treatment, e.g., based on measuring high TMB.

[0287] Antibodies that specifically bind to KIR block the interaction between killer cell immunoglobulin-like receptors (KIR) and their ligands on NK cells. Blocking these receptors promotes NK cell activation and, potentially, tumor cell destruction by the latter. Examples of anti-KIR antibodies are disclosed in International Publications WO / 2014 / 055648, WO2005 / 003168, WO2005 / 009465, WO2006 / 072625, WO2006 / 072626, WO2007 / 042573, WO2008 / 084106, WO2010 / 065939, WO2012 / 071411, and WO / 2012 / 160448.

[0288] One anti-KIR antibody useful in the present invention is lirilumab (also referred to as BMS-986015, IPH2102, or the S241P variant of 1-7F9), first described in International Publication WO2008 / 084106. An additional anti-KIR antibody useful in the present invention is 1-7F9 (also referred to as IPH2101), described in International Publication WO2006 / 003179. In certain embodiments, the anti-KIR antibody for the present compositions cross-competes with lirilumab or I-7F9 for binding to KIR. In other embodiments, the anti-KIR antibody binds to the same epitope as lirilumab or I-7F9. In other embodiments, the anti-KIR antibody comprises the six CDRs of lirilumab or I-7F9.

[0289] Anti-GITR antibody

[0006] Certain aspects of the present invention relate to methods of treating a subject with a tumor having an elevated TMB status, comprising administering to the subject an immunotherapeutic agent, wherein the immunotherapeutic agent comprises an anti-GITR antibody or antigen-binding portion thereof. The method may further comprise measuring the TMB status of a biological sample from the subject. Furthermore, the present invention contemplates administering the anti-GITR antibody or antigen-binding portion thereof to a subject identified as suitable for such treatment, e.g., based on measuring elevated TMB.

[0290] The anti-GITR antibody can be any anti-GITR antibody that specifically binds to the human GITR target and activates the glucocorticoid-induced tumor necrosis factor receptor (GITR). GITR is a member of the TNF receptor superfamily that is expressed on the surface of multiple types of immune cells, including regulatory T cells, effector T cells, B cells, natural killer (NK) cells, and activated dendritic cells ("anti-GITR agonist antibodies"). In particular, GITR activation increases the proliferation and function of effector T cells and abolishes the suppression by activated T regulatory cells. Furthermore, GITR stimulation promotes anti-tumor immunity by increasing the activity of other immune cells, such as NK cells, antigen-presenting cells, and B cells. Examples of anti-GITR antibodies are disclosed in International Publications WO / 2015 / 031667, WO2015 / 184,099, WO2015 / 026,684, WO11 / 028683, and WO / 2006 / 105021, U.S. Patents 7,812,135 and 8,388,967, and U.S. Publications 2009 / 0136494, 2014 / 0220002, 2013 / 0183321, and 2014 / 0348841.

[0291] In one embodiment, the anti-GITR antibody useful in the present invention is TRX518 (e.g., as described in Schaer et al. Curr Opin Immunol. (2012) Apr;24(2):217-224 and WO / 2006 / 105021). In another embodiment, the anti-GITR antibody is selected from MK4166, MK1248, and the antibodies described in WO11 / 028683 and U.S. Pat. No. 8,709,424, e.g., comprising a VH chain comprising SEQ ID NO: 104 and a VL chain comprising SEQ ID NO: 105 (wherein the SEQ ID NOs are based on WO11 / 028683 or U.S. Pat. No. 8,709,424). In certain embodiments, the anti-GITR antibody is an anti-GITR antibody disclosed in WO2015 / 031667, e.g., an antibody comprising VH CDRs 1-3 comprising SEQ ID NOs: 31, 71, and 63, respectively, of WO2015 / 031667, and VL CDRs 1-3 comprising SEQ ID NOs: 5, 14, and 30, respectively, of WO2015 / 031667. In certain embodiments, the anti-GITR antibody is an anti-GITR antibody disclosed in WO2015 / 184099, e.g., antibody Hum231#1 or Hum231#2, or a CDR or derivative thereof (e.g., pab1967, pab1975, or pab1979). In some embodiments, the anti-GITR antibody is an anti-GITR antibody disclosed in JP2008278814, WO09 / 009116, WO2013 / 039954, US20140072566, US20140072565, US20140065152, or WO2015 / 026684, or is INBRX-110 (INHIBRx), LKZ-145 (Novartis), or MEDI-1873 (MedImmune). In some embodiments, the anti-GITR antibody is an anti-GITR antibody described in PCT / US2015 / 033991 (e.g., an antibody comprising the variable regions of 28F3, 18E10, or 19D3). For example, the anti-GITR antibody may be an antibody comprising the following VH and VL chains or their CDRs:

[0292] VH: QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVIWYEGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGGSMVRGDYYYGMDVWGQGTTVTVS (SEQ ID NO: 1) and

[0293] VL: AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSYPYTFGQGTKLEIK (SEQ ID NO: 2); or

[0294] VH: QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGFHWVRQAPGKGLEWVAVIWYAGSNKFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGGQLDYYYYYVMDVWGQGTTVTVSS (SEQ ID NO: 3) and

[0295] VL: DIQMTQSPSSLSASVGDRVTITCRASQGISSWLAWYQQKPEKAPKSLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSYPYTFGQGTKLEIK (SEQ ID NO: 4); or

[0296] VH: VQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVIWYAGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGGRIAVAFYYSMDVWGQGTTVTVSS (SEQ ID NO: 5) and

[0297] VL: DIQMTQSPSSLSASVGDRVTITCRASQGISSWLAWYQQKPEKAPKSLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSYPYTFGQGTKLEIK (SEQ ID NO: 6).

[0298] In some embodiments, the antibody comprising the above-described VH heavy chain and VL light chain or their CDRs comprises a heavy chain constant region of the IgG1 isotype that is wild-type or mutated, e.g., to render it effector-less. In some embodiments, the anti-GITR antibody comprises the following heavy and light chain amino acid sequences:

[0299] Heavy chain: QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVIWYEGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGGSMVRGDYYYGMDVW GQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVE CPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 7) and

[0300] Light chain: AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSYPYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 8) or

[0301] Heavy chain: qvqlvesgggvvqpgrslrlscaasgftfssygmhwvrqapgkglewvaviwyegsnkyyadsvkgrftisrdnskntlylqmnslraedtavyycarggsmvrgdyyygmdvwg qgttvtvssastkgpsvfplapsskstsggtaalgclvkdyfpepvtvswnsgaltsgvhtfpavlqssglyslssvvtvpssslgtqtyicnvnhkpsntkvdkrvepkscdkth tcppcpapeaegapsvflfppkpkdtlmisrtpevtcvvvdvshedpevkfnwyvdgvevhnaktkpreeqynstyrvvsvltvlhqdwlngkeykckvsnkalpssiektiskakgqprepqvytlppsreemtknqvsltclvkgfypsdiavewesngqpennykttppvldsdgsfflyskltvdksrwqqgnvfscsvmhealhnhytqkslslspg (SEQ ID NO: 9) and

[0302] Light chain: AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSYPYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 10).

[0303] In some embodiments, the anti-GITR antibody cross-competes with an anti-GITR antibody described herein, e.g., TRX518, MK4166, or an antibody comprising the VH and VL domain amino acid sequences described herein. In some embodiments, the anti-GITR antibody binds to the same epitope as an anti-GITR antibody described herein, e.g., TRX518, MK4166, or an antibody comprising the VH and VL domain amino acid sequences described herein. In some embodiments, the anti-GITR antibody comprises the six CDRs of TRX518, MK4166, or an antibody comprising the VH and VL domain amino acid sequences described herein.

[0304] Further antibodies In certain embodiments, the immunotherapeutic agent comprises an anti-TGFβ antibody. In certain embodiments, the anti-TGFβ antibody is an anti-TGFβ antibody disclosed in International Publication WO / 2009 / 073533.

[0305] In certain embodiments, the immunotherapeutic agent comprises an anti-IL-10 antibody. In certain embodiments, the anti-IL-10 antibody is an anti-IL-10 antibody disclosed in International Publication WO / 2009 / 073533.

[0306] In certain other embodiments, the immunotherapeutic agent comprises an anti-B7-H4 antibody. In certain embodiments, the anti-B7-H4 antibody is the anti-B7-H4 antibody disclosed in International Publication WO / 2009 / 073533.

[0307] In certain embodiments, the immunotherapeutic agent comprises an anti-Fas ligand antibody. In certain embodiments, the anti-Fas ligand antibody is an anti-Fas ligand antibody disclosed in International Publication WO / 2009 / 073533.

[0308] In some embodiments, the immunotherapeutic agent comprises an anti-CXCR4 antibody. In some embodiments, the anti-CXCR4 antibody is an anti-CXCR4 antibody disclosed in U.S. Publication No. 2014 / 0322208 (e.g., urocuplumab (BMS-936564)).

[0309] In some embodiments, the immunotherapeutic agent comprises an anti-mesothelin antibody, ie, the anti-mesothelin antibody is an anti-mesothelin antibody disclosed in U.S. Patent No. 8,399,623.

[0310] In some embodiments, the immunotherapeutic agent comprises an anti-HER2 antibody. In some embodiments, the anti-HER2 antibody is Herceptin (U.S. Patent No. 5,821,337), trastuzumab, or ado-trastuzumab emtansine (Kadcyla, e.g., WO / 2001 / 000244).

[0311] In some embodiments, the immunotherapeutic agent comprises an anti-CD27 antibody. In some embodiments, the anti-CD27 antibody is varlilumab (also known as "CDX-1127" and "1F5"), a human IgG1 antibody that is an agonist of human CD27, e.g., as disclosed in U.S. Pat. No. 9,169,325.

[0312] In certain embodiments, the immunotherapeutic agent comprises an anti-CD73 antibody. In certain embodiments, the anti-CD73 antibody is CD73.4.IgG2C219S.IgG1.1f.

[0313] In some embodiments, the immunotherapeutic agent comprises an anti-MICA antibody. As used herein, an anti-MICA antibody is an antibody or antigen-binding fragment thereof that specifically binds to MHC class I polypeptide-related sequence A. In some embodiments, the anti-MICA antibody binds to MICB in addition to MICA. In some embodiments, the anti-MICA antibody inhibits membrane-bound MICA and soluble MICA release. In some embodiments, the anti-MICA antibody is an anti-MICA antibody disclosed in U.S. Publication No. 2014 / 004112A1, U.S. Publication No. 2016 / 046716A1, or U.S. Publication No. 2017 / 022275A1.

[0314] In some embodiments, the immunotherapeutic agent comprises an anti-TIM3 antibody. As used herein, the anti-TIM3 antibody is an antibody or an antigen-binding fragment thereof that specifically binds to T-cell immunoglobulin and mucin domain-containing-3 (TIM3), also known as hepatitis A virus cellular receptor 2 (HAVCR2). In some embodiments, the anti-TIM3 antibody can stimulate an immune response, for example, an antigen-specific T cell response. In some embodiments, the anti-TIM3 antibody binds to soluble or membrane-bound human or cyno TIM3. In some embodiments, the anti-TIM3 antibody is the anti-TIM3 antibody disclosed in International Publication WO / 2018 / 013818, the entire contents of which are incorporated herein by reference.

[0315] Standard treatment for SCLC In some embodiments, the subject has been previously treated with one or more standard therapies for SCLC. In some embodiments, the subject is refractory to one or more standard therapies for SCLC. In some embodiments, the subject shows progressive disease after one or more standard therapies for SCLC. In some embodiments, the subject shows stable disease after one or more standard therapies for SCLC. In some embodiments, the subject is not refractory to one or more standard therapies for SCLC. In some embodiments, the subject has relapsed after one or more standard therapies for SCLC.

[0316] Standard treatments for various types of cancer are well known to those skilled in the art. For example, the National Comprehensive Cancer Network (NCCN), an alliance of 21 major cancer centers in the United States, publishes the NCCN Clinical Practice Guidelines in Oncology (NCCN Guidelines®), which provide detailed, state-of-the-art information on standard treatments for a wide variety of cancers (see NCCN Guidelines® (2014), available at http: / / www.nccn.org / professionals / physician_gls / f_guidelines.asp (last accessed June 2, 2016)).

[0317] Surgery, radiation therapy (RT), and chemotherapy are three modalities commonly used to treat patients with SCLC. The most commonly used initial chemotherapy regimen is etoposide (TOPOSAR® or Bepcid®) in combination with cisplatin (Platinol®), known as EP. For people with extensive-stage small cell lung cancer, chemotherapy alone using the EP regimen is the standard treatment. However, another regimen that may be used is carboplatin (Paraplatin®) in combination with irinotecan (CAMPTOSAR®).

[0318] Although SCLC is highly sensitive to initial treatments including chemotherapy and / or radiation therapy, the majority of patients ultimately die from recurrence of SCLC. Therefore, there is a particular unmet need in patients with recurrent SCLC due to the lack of effective treatments after first-line therapy.

[0319] Pharmaceutical Compositions and Dosages Therapeutic agents of the present invention can be compositions, such as pharmaceutical compositions comprising an antibody and / or cytokine and a pharmaceutically acceptable carrier. As used herein, "pharmaceutically acceptable carrier" includes any and all physiologically compatible solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. In some embodiments, the carrier for an antibody-containing composition is suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal, or epidermal administration (e.g., by injection or infusion), while the carrier for an antibody and / or cytokine-containing composition is suitable for non-parenteral administration, e.g., oral administration. In some embodiments, subcutaneous injection is based on Halozyme Therapeutics' ENHANZE® drug delivery technology (see U.S. Pat. No. 7,767,429, incorporated herein by reference in its entirety). ENHANZE® is a co-formulation of an antibody and a recombinant human hyaluronidase enzyme (rHuPH20), which eliminates the traditional volumetric constraints of biologics and drugs through the extracellular matrix, enabling subcutaneous delivery (see U.S. Pat. No. 7,767,429). Pharmaceutical compositions of the present invention may contain one or more pharmaceutically acceptable salts, antioxidants, aqueous and non-aqueous carriers, and / or adjuvants, such as preservatives, wetting agents, emulsifying agents, and dispersing agents. Thus, in some embodiments, pharmaceutical compositions of the present invention further comprise a recombinant human hyaluronidase enzyme, such as rHuPH20.

[0320] Dosing regimens are adjusted to provide the optimal desired response, e.g., maximal therapeutic response and / or minimal adverse effects. In certain embodiments, the anti-PD-1 antibody, anti-PD-L1 antibody, and / or anti-CTLA-4 antibody are administered at a weight-based dose. For administration of an anti-PD-1 antibody as monotherapy or in combination with other anti-cancer agents (e.g., in combination with an anti-CTLA-4 antibody), the dosage may be in the range of about 0.01 to about 20 mg / kg, about 0.1 to about 10 mg / kg, about 0.1 to about 5 mg / kg, about 0.01 mg / kg to about 5 mg / kg, about 1 to about 5 mg / kg, about 2 to about 5 mg / kg, about 1 to about 3 mg / kg, about 7.5 to about 12.5 mg / kg, or about 0.1 to about 30 mg / kg of the subject's body weight. For example, the dosage can be about 0.1 mg / kg body weight, about 0.3 mg / kg body weight, about 1 mg / kg body weight, about 2 mg / kg body weight, about 3 mg / kg body weight, about 5 mg / kg body weight, or about 10 mg / kg body weight, and more preferably 0.3 mg / kg body weight, 1 mg / kg body weight, 2 mg / kg body weight, 3 mg / kg body weight, or 5 mg / kg body weight. In one embodiment, the dosage of the anti-PD-1 antibody is 3 mg / kg body weight. The administration schedule is generally based on the typical pharmacokinetics of the antibody and is designed to achieve an exposure that results in sustained receptor occupancy (RO). Exemplary treatment regimens include administration about once weekly, about every 2 weeks, about every 3 weeks, about every 4 weeks, about once a month, about every 3 to 6 months, or at longer intervals. In one embodiment, an anti-PD-1 antibody, such as nivolumab, is administered to a subject about every 2 weeks. In another embodiment, the antibody is administered about every 3 weeks. The dosage and schedule can vary during treatment. For example, dosing schedules for anti-PD-1 monotherapy can include administration of the antibody (i) about every 2 weeks in an about 6-week cycle; (ii) about every 4 weeks for about 6 doses, followed by about every 3 months; (iii) about every 3 weeks; or (iv) about 3 mg / kg to about 10 mg / kg once, followed by about 1 mg / kg about every 2-3 weeks, taking into account that IgG4 antibodies generally have a half-life of 2-3 weeks.

[0321] In certain embodiments, the dosing regimen for an anti-PD-1 antibody or anti-PD-L1 antibody of the invention comprises intravenous administration of about 0.3-1 mg / kg body weight, about 5 mg / kg body weight, 1-5 mg / kg body weight, or about 1 to about 3 mg / kg body weight, where the antibody is administered about every 14 to 21 days in up to about 6-week or about 12-week cycles until complete response or confirmed disease progression. In certain embodiments, anti-PD-1 antibody monotherapy is administered at 3 mg / kg every 2 weeks until disease progression or unacceptable toxicity. In certain embodiments, the antibody treatment or any combination treatment disclosed herein is continued for at least about 1 month, at least about 3 months, at least about 6 months, at least about 9 months, at least about 1 year, at least about 18 months, at least about 24 months, at least about 3 years, at least about 5 years, or at least about 10 years.

[0322] When used in combination with other cancer drugs (e.g., in combination with an anti-CTLA-4 antibody), the dosage of the anti-PD-1 antibody may be reduced compared to the monotherapy dose. A dose of nivolumab typically lower than 3 mg / kg but greater than 0.001 mg / kg is a subtherapeutic dose. A subtherapeutic dose of the anti-PD-1 antibody used in the methods herein is greater than 0.001 mg / kg and less than 3 mg / kg. In certain embodiments, the subtherapeutic dose is about 0.001 mg / kg to about 1 mg / kg, about 0.01 mg / kg to about 1 mg / kg, about 0.1 mg / kg to about 1 mg / kg, or about 0.001 mg / kg to about 0.1 mg / kg of body weight. In certain embodiments, the subtherapeutic dose is at least about 0.001 mg / kg, at least about 0.005 mg / kg, at least about 0.01 mg / kg, at least about 0.05 mg / kg, at least about 0.1 mg / kg, at least about 0.5 mg / kg, or at least about 1.0 mg / kg body weight. Receptor occupancy data from 15 subjects receiving nivolumab at doses of 0.3 mg / kg to 10 mg / kg indicate that PD-1 occupancy appears to be dose-independent within this dose range. Across all doses, the mean occupancy was 85% (range, 70%-97%), and the mean plateau occupancy was 72% (range, 59%-81%). In certain embodiments, a 0.3 mg / kg dose may allow sufficient exposure to result in maximal biological activity. Receptor occupancy data from 15 subjects receiving nivolumab at doses of 0.3 mg / kg to 10 mg / kg indicate that PD-1 occupancy appears to be dose-independent within this dose range. Across all doses, the mean occupancy was 85% (range, 70%-97%), with a mean plateau occupancy of 72% (range, 59%-81%) (Brahmer et al., J Clin Oncol 28:3167-75 2010). Thus, a 0.3 mg / kg dose may provide sufficient exposure to produce maximal bioactivity.

[0323] Although high nivolumab monotherapy doses of up to 10 mg / kg every 2 weeks have been achieved without reaching the maximum tolerated dose (MTD), the substantial toxicity reported in other trials of checkpoint inhibitor and antiangiogenic therapy combinations (see, e.g., Johnson et al. (2013) Cancer Immunol Res 1:373-77; Rini et al. (2011) Cancer 117:758-67) supports the selection of nivolumab doses below 10 mg / kg.

[0324] In certain embodiments, the dose of the anti-PD-1 antibody (or anti-PD-L1 antibody) is a fixed dose in the pharmaceutical composition. In other embodiments, the methods of the present invention can be used with fixed doses (doses given to patients regardless of the patient's weight). For example, the fixed dose of nivolumab can be about 240 mg. For example, the fixed dose of pembrolizumab can be about 200 mg. In certain embodiments, the anti-PD-1 antibody is administered at a dose of about 240 mg. In certain embodiments, the anti-PD-1 antibody is administered at a dose of about 360 mg. In certain embodiments, the anti-PD-1 antibody is administered at a dose of about 480 mg. In certain embodiments, 360 mg of the anti-PD-1 antibody is administered every three weeks. In other embodiments, 480 mg of the anti-PD-1 antibody is administered every four weeks.

[0325] Ipilimumab (Yervoy®) is approved for the treatment of melanoma at 3 mg / kg intravenously every three weeks for four doses. Thus, in certain embodiments, about 3 mg / kg is the maximum dose of ipilimumab used in combination with an anti-PD-1 antibody, but in certain embodiments, when combined with nivolumab, an anti-CTLA-4 antibody such as ipilimumab may be administered within the range of about 0.3 to about 10 mg / kg, about 0.5 to about 10 mg / kg, about 0.5 to about 5 mg / kg, or about 1 to about 5 mg / kg body weight about every two or three weeks. In other embodiments, ipilimumab is administered on a different dosing schedule than nivolumab. In some embodiments, ipilimumab is administered about weekly, about every 2 weeks, about every 3 weeks, about every 4 weeks, about every 5 weeks, about every 6 weeks, about every 7 weeks, about every 8 weeks, about every 9 weeks, about every 10 weeks, about every 11 weeks, about every 12 weeks, or about every 15 weeks. Typically, a dose of ipilimumab less than 3 mg / kg every 3 weeks but greater than 0.001 mg / kg is a subtherapeutic dose. A subtherapeutic dose of an anti-CTLA-4 antibody used in the methods herein is greater than 0.001 mg / kg and less than 3 mg / kg. In some embodiments, a subtherapeutic dose is between about 0.001 mg / kg and about 1 mg / kg, between about 0.01 mg / kg and about 1 mg / kg, between about 0.1 mg / kg and about 1 mg / kg, or between about 0.001 mg / kg and about 0.1 mg / kg of body weight. In some embodiments, the subtherapeutic dose is at least about 0.001 mg / kg, at least about 0.005 mg / kg, at least about 0.01 mg / kg, at least about 0.05 mg / kg, at least about 0.1 mg / kg, at least about 0.5 mg / kg, or at least about 1.0 mg / kg body weight. It has been shown that the combination of nivolumab 3 mg / kg and ipilimumab 3 mg / kg exceeds the MTD in melanoma patients, but the combination of nivolumab 1 mg / kg and ipilimumab 3 mg / kg or nivolumab 3 mg / kg and ipilimumab 1 mg / kg has been found to be tolerable in melanoma patients (Wolchok et al., N Engl JMed 369(2):122-33(2013)).Thus, nivolumab is tolerated at up to 10 mg / kg intravenously every two weeks, although in some embodiments, when combined with ipilimumab, the dose of the anti-PD-1 antibody does not exceed about 3 mg / kg. In some embodiments, based on risk-benefit and PK-PD assessment, the dosages used include combinations of about 1 mg / kg nivolumab and about 3 mg / kg ipilimumab, about 3 mg / kg nivolumab and about 1 mg / kg ipilimumab, or about 3 mg / kg nivolumab and about 3 mg / kg ipilimumab, each administered at a dosing frequency of about every 2 to 4 weeks, in some embodiments, about every 2 weeks or about every 3 weeks. In certain other embodiments, nivolumab is administered at a dosage of about 0.1 mg / kg, about 0.3 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, or about 5 mg / kg, in combination with ipilimumab administered at a dosage of about 0.1 mg / kg, about 0.3 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, or about 5 mg / kg about every 2 weeks, about every 3 weeks, or about every 4 weeks.

[0326] In one embodiment, a combination of an anti-PD-1 antibody and an anti-CTLA-4 antibody is administered intravenously to a subject in the induction phase once, twice, three times, or four times approximately every two or three weeks. In one embodiment, a combination of nivolumab and ipilimumab is administered intravenously to a subject in the induction phase approximately four times every two or three weeks. The induction phase is followed by a maintenance phase, during which the anti-PD-1 antibody alone is administered to the subject at a dose of about 0.1 mg / kg, about 0.3 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 5 mg / kg, or about 10 mg / kg approximately every two or three weeks, as long as treatment is shown to be effective or until unmanageable toxicity or disease progression occurs. In one embodiment, nivolumab is administered at a dose of about 3 mg / kg body weight approximately every two weeks during the maintenance phase. In one embodiment, the anti-PD-1 antibody is administered at a dose of about 3 mg / kg (or a flat dose of 240 mg) about every two weeks, and the anti-CTLA-4 antibody is administered at a dose of 1 mg / kg about every six weeks. In one embodiment, the anti-PD-1 antibody is administered at a flat dose (e.g., 240 mg or 480 mg) and the anti-CTLA-4 antibody is administered at a weight-based dose (e.g., 1 mg / kg or 3 mg / kg).

[0327] The antibodies disclosed herein can be administered according to a "treatment cycle" or "cycle" (these terms are used interchangeably herein). As used herein, the term "cycle" refers to a series of treatments repeated on a regular schedule with rest periods in between. For example, a treatment administered for one week followed by a three-week rest period is a treatment cycle. In certain embodiments, the anti-PD-1 antibody and / or anti-CTLA-4 antibody are administered for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 cycles. In certain embodiments, the administration of the anti-PD-1 antibody and anti-CTLA-4 antibody is repeated four times (4 cycles).

[0328] In certain embodiments, the anti-PD-1 antibody (or anti-CTLA-4 antibody) and anti-CTLA-4 antibody are formulated as a single composition, wherein the dose of the anti-PD1 antibody and the dose of the anti-CTLA-4 antibody are combined in a ratio of 1:50, 1:40, 1:30, 1:20, 1:10, 1:5, 1:3, 1:1, 3:1, 5:1, 10:1, 20:1, 30:1, 40:1, or 50:1. In other embodiments, the dose of the anti-CTLA-4 antibody is a fixed dose. In some embodiments, the ratio of anti-PD-1 antibody or anti-PD-L1 antibody to anti-CTLA-4 antibody is at least about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, about 1:30, about 1:40, about 1:50, about 1:60, about 1:70, about 1:80, about 1:90, about 1:100, about 1:120, about 1:1 1:40, about 1:160, about 1:180, about 1:200, about 200:1, about 180:1, about 160:1, about 140:1, about 120:1, about 100:1, about 90:1, about 80:1, about 70:1, about 60:1, about 50:1, about 40:1, about 30:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1 or about 2:1.

[0329] In some embodiments, the dose of the anti-CTLA-4 antibody is a fixed dose administered to patients regardless of body weight. In certain embodiments, the fixed dose of the anti-CTLA-4 antibody is about 80 mg. In some embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody is administered as a fixed dose monotherapy. In some embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody is administered as a fixed dose in combination with the anti-CTLA-4 antibody. In some embodiments, the fixed dose of the anti-PD-1 antibody or anti-PD-L1 antibody is at least about 100-600 mg, e.g., at least about 200-300 mg, at least about 400-500 mg, or at least about 240 mg or at least about 480 mg, such as at least about 60 mg, at least about 80 mg, at least about 100 mg, at least about 120 mg, at least about 140 mg, at least about 160 mg, at least about 180 mg, at least about 200 mg, at least about 220 mg, at least about 240 mg, at least about 260 mg, at least about 280 mg, at least about 320 mg, at least about 360 mg, at least about 400 mg, at least about 440 mg, at least about 480 mg, at least about 520 mg, at least about 560 mg, at least about 600 mg, or at least about 660 mg, or at least about 720 mg. In some embodiments, the fixed dose of the anti-PD-1 antibody or anti-PD-L1 antibody is at least about 600 to 1200 mg. In some embodiments, the fixed dose of the anti-PD-1 antibody or anti-PD-L1 antibody is at least about 600 mg, at least about 640 mg, at least about 680 mg, at least about 720 mg, at least about 760 mg, at least about 800 mg, at least about 840 mg, at least about 880 mg, at least about 920 mg, at least about 960 mg, at least about 1000 mg, at least about 1040 mg, at least about 1080 mg, at least about 1120 mg, at least about 1160 mg, or at least about 1200 mg. In some embodiments, the anti-PD-1 antibody or antigen-binding portion thereof is administered at a dose of at least about 240 mg or at least about 480 mg about every 2 weeks or 4 weeks.In certain embodiments, the anti-PD-L1 antibody or antigen-binding portion thereof is administered at a dose of at least about 240 mg or at least about 480 mg about every two weeks or four weeks. In certain embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody is administered at a dose of at least about 720 mg. In certain embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody is administered at a dose of at least about 960 mg. In certain embodiments, the anti-PD-1 antibody or anti-PD-L1 antibody is administered at a dose of at least about 1200 mg.

[0330] In other embodiments, the anti-PD-1 antibody or its antigen-binding portion is administered at a dose of greater than about 240 mg, i.e., at least about 240 mg. When used in combination with other anti-cancer drugs, the dose of the anti-PD-1 antibody can be reduced compared to the monotherapy dose. For example, a dose significantly lower than the typical 3 mg / kg every 3 weeks of nivolumab, such as 0.1 mg / kg, or less frequently than every 3 or 4 weeks, is considered a subtherapeutic dose.

[0331] In one embodiment, the fixed doses of the anti-PD-1 antibody and anti-CTLA-4 antibody in the combination are about 60 mg to about 80 mg for the anti-PD-1 antibody, e.g., about 60 mg, about 65 mg, about 70 mg, about 75 mg, or about 80 mg, and about 180 mg to about 240 mg for the anti-CTLA-4 antibody, e.g., about 180 mg, about 190 mg, about 200 mg, about 210 mg, about 220 mg, about 230 mg, or about 240 mg. In another embodiment, the fixed doses of the anti-PD-1 antibody and anti-CTLA-4 antibody in the combination are about 80 mg for the anti-PD-1 antibody and about 240 mg for the anti-CTLA-4 antibody. The anti-PD-1 antibody and anti-CTLA-4 antibody may be in the same formulation or separate formulations.

[0332] For combinations of nivolumab with other anti-cancer agents, these agents are preferably administered at their approved dosages. Treatment continues as long as clinical benefit is observed or until unacceptable toxicity or disease progression occurs. Nevertheless, in certain embodiments, the dosages of these anti-cancer agents administered are significantly lower than their approved dosages, i.e., subtherapeutic dosages of the agents are administered in combination with anti-PD-1 antibodies or anti-PD-L1 antibodies. The anti-PD-1 antibodies or anti-PD-L1 antibodies may be administered at dosages shown in clinical trials to produce maximal efficacy as monotherapy, for example, nivolumab at about 3 mg / kg every 3 weeks (Topalian et al., N Engl JMed 366:2443-54 (2012a); Topalian et al., Curr Opin Immunol 24:207-12 (2012b)), or at significantly lower dosages, i.e., subtherapeutic dosages. In one embodiment, the anti-PD-1 antibody is administered at about 3 mg / kg about every 3 weeks.

[0333] The dosage and frequency vary depending on the half-life of the antibody in the subject. Generally, human antibodies have the longest half-life, followed by humanized antibodies, chimeric antibodies, and non-human antibodies. The dosage and frequency can vary depending on whether the treatment is preventive or therapeutic. In preventive applications, relatively low dosages are generally administered at relatively infrequent intervals over a long period of time. Some patients continue to receive treatment for the rest of their lives. In therapeutic applications, relatively high dosages at relatively short intervals may be required until the progression of the disease is reduced or terminated, preferably until the patient shows partial or complete improvement in disease symptoms. Thereafter, the patient can be administered a preventive regimen.

[0334] In some embodiments, a subject treated with a combination of an anti-PD-1 antibody and an anti-CTLA-4 antibody may be further treated with anti-PD-1 antibody monotherapy. In some embodiments, the subject is treated with a fixed dose of an anti-PD-1 antibody and an anti-CTLA-4 antibody in combination, where the anti-PD-1 antibody is about 60 mg to about 80 mg, e.g., about 60 mg, about 65 mg, about 70 mg, about 75 mg, or about 80 mg, and the anti-CTLA-4 antibody is about 180 mg to about 240 mg, e.g., about 180 mg, about 190 mg, about 200 mg, about 210 mg, about 220 mg, about 230 mg, or about 240 mg, followed by anti-PD-1 monotherapy, e.g., about 3 mg / kg or about 60 to about 80 mg, e.g., about 60 mg, about 65 mg, about 70 mg, about 75 mg, or about 80 mg. In other embodiments, the subject is treated with fixed doses of an anti-PD-1 antibody and an anti-CTLA-4 antibody in combination, which is about 80 mg for the anti-PD-1 and about 240 mg for the anti-CTLA-4 antibody, followed by an anti-PD-1 antibody monotherapy, about 3 mg / kg or about 60 mg to about 80 mg, e.g., about 60 mg, about 65 mg, about 70 mg, about 75 mg, or about 80 mg.

[0335] The actual dosage level of the active ingredient in the pharmaceutical composition of the present invention can be varied to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and administration method without causing undue toxicity to the patient. The selected dosage level depends on various pharmacokinetic factors, including the activity of the particular composition of the present invention used, the route of administration, the time of administration, the excretion rate of the particular compound used, the duration of treatment, other drugs, compounds and / or substances used in combination with the particular composition used, the age, sex, weight, condition, general health and medical history of the patient being treated, and similar factors well known in the pharmaceutical arts. The compositions of the present invention can be administered by one or more routes of administration using one or more of a variety of methods well known in the art. As will be recognized by those skilled in the art, the route and / or method of administration will depend on the desired results.

[0336] kit Also within the scope of the present invention are kits containing anti-PD-1 antibodies and / or anti-CTLA-4 antibodies for therapeutic use. The kits generally include a label containing the intended use and instructions for use of the kit's contents. The term label includes any writing or recording medium provided on or with the kit or that otherwise accompanies the kit. Accordingly, the present invention provides kits for treating a subject with a tumor, the kit comprising: (a) an antibody or antigen-binding portion thereof that specifically binds to the PD-1 receptor and inhibits PD-1 activity ("anti-PD-1 antibody"), at a dose ranging from 0.1 to 10 mg / kg body weight; and (b) instructions for using the anti-PD-1 antibody in the methods disclosed herein. The present invention further provides kits for treating a subject with an SCLC-derived tumor, the kit comprising: (a) an anti-PD-1 antibody at a dose ranging from 0.1 to 10 mg / kg body weight; (b) an anti-CTLA-4 antibody at a dose ranging from 0.1 to 10 mg / kg body weight; and (c) instructions for using the anti-PD-1 antibody and anti-CTLA-4 antibody in the methods disclosed herein. In certain embodiments, the present invention provides a kit for treating a subject with a tumor derived from SCLC, the kit comprising: (a) an anti-PD-1 antibody at a dose ranging from 200 mg to 800 mg or an anti-PD-L1 antibody at a dose ranging from 200 mg to 1800 mg; (b) instructions for using the anti-PD-1 antibody or anti-PD-L1 antibody in a method disclosed herein. In certain embodiments, the present invention provides a kit for treating a subject with a tumor derived from SCLC, the kit comprising: (a) an anti-PD-1 antibody at a dose ranging from 200 mg to 800 mg or an anti-PD-L1 antibody at a dose ranging from 200 mg to 1800 mg; (b) an anti-CTLA-4 antibody at a dose ranging from 80 mg to 800 mg; and (c) instructions for using (a) the anti-PD-1 antibody or anti-PD-L1 antibody and (b) the anti-CTLA-4 antibody in a method disclosed herein. In certain embodiments, the tumor is lung cancer, e.g., SCLC.

[0337] In a preferred embodiment, for the treatment of a human patient, the kit comprises an anti-human PD-1 antibody disclosed herein, such as nivolumab or pembrolizumab. In a preferred embodiment, for the treatment of a human patient, the kit comprises an anti-human PD-L1 antibody disclosed herein, such as atezolizumab, durvalumab, or avelumab. In a preferred embodiment, for the treatment of a human patient, the kit comprises an anti-human CTLA-4 antibody disclosed herein, such as ipilimumab, tremelimumab, MK-1308, or AGEN-1884.

[0338] In some embodiments, the kit further comprises a cytokine or a variant thereof. In some embodiments, the kit comprises (a) an anti-PD-1 antibody or an anti-PD-L1 antibody, (b) an anti-CTLA-4 antibody, and (c) a CD122 agonist.

[0339] In some embodiments, the kit further comprises a comprehensive genomic profiling assay as disclosed herein. In some embodiments, the kit comprises FOUNDATIONONE® CDX TM and a genomic profiling assay. In certain embodiments, the kit further comprises instructions for administering an immunotherapeutic agent, e.g., an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-CTLA-4 antibody, and / or a cytokine, to a subject identified as having a high TMB status by the methods disclosed herein. In other embodiments, the kit further comprises instructions for administering (a) an anti-PD-1 antibody or an anti-PD-L1 antibody, (b) an anti-CTLA-4 antibody, and (c) a cytokine, e.g., a CD122 agonist, to a subject identified as having a high TMB status by the methods disclosed herein.

[0340] All references cited above, as well as all references cited herein, are incorporated by reference in their entirety.

[0341] The following examples are offered by way of illustration and not by way of limitation. [Example]

[0342] Example 1. Patients with recurrent SCLC have limited treatment options and poor survival rates. Early results from clinical trials of patients with SCLC treated with nivolumab alone or in combination with ipilimumab demonstrated durable responses and encouraging survival. 26% of patients receiving the combination of nivolumab and ipilimumab had an overall survival rate of more than 2 years, compared with 14% of patients receiving nivolumab monotherapy. These data supported the inclusion of nivolumab, with or without ipilimumab, in the NCCN Guidelines for the treatment of SCLC.

[0343] PD-L1 expression on tumor cells is rare in SCLC, and responses have been observed regardless of PD-L1 status. Improved biomarkers are needed for immunotherapy of SCLC. Previously, subjects with high TMB were found to have higher rates of progression-free survival (PFS) after treatment with nivolumab monotherapy compared with subjects with low or intermediate TMB. SCLC is seen almost exclusively in patients with a smoking history and is characterized by high TMB. Correlations between TMB and efficacy have been seen with nivolumab in NSCLC and bladder cancer and with ipilimumab in melanoma. High TMB may be associated with increased benefit from nivolumab with or without ipilimumab in SCLC. This clinical trial will investigate the use of tumor mutation burden (TMB) as a predictive biomarker for nivolumab with or without ipilimumab in SCLC.

[0344] Clinical trial design Subjects were selected who had a prior diagnosis of SCLC and had received at least one prior platinum-containing regimen (Figure 23). Non-randomized and randomized (3:2) patients received either (1) nivolumab monotherapy, including nivolumab 3 mg / kg IV every 2 weeks, until disease progression or unacceptable toxicity; or (2) nivolumab / ipilimumab combination therapy, including nivolumab 1 mg / kg and ipilimumab 3 mg / kg IV every 3 weeks, for four cycles, followed by nivolumab monotherapy, including nivolumab 3 mg / kg IV every 2 weeks, until disease progression or unacceptable toxicity.

[0345] The primary objective was to measure the objective response rate (ORR) by RECIST v1.1. Secondary objectives included monitoring safety, overall survival (OS), progression-free survival (PFS), and duration of response (DOR). Prespecified study objectives included biomarker analysis and health status using the EQ-5D instrument.

[0346] TMB was determined by whole-exome sequencing using an Illumina HiSeq 2500 with 2 × 100 bp paired-end reads and calculated as the total number of nonsynonymous missense mutations in the tumor. For exploratory analyses, patients were divided into three subgroups based on TMB tertiles.

[0347] Baseline A total of 245 subjects were included in the nivolumab monotherapy (ITT) study, of which 133 were TMB-evaluable (Table 15 and Figure 2). A total of 156 subjects were included in the nivolumab / ipilimumab combination therapy (ITT) study, of which 78 were TMB-evaluable (Table 15 and Figure 2). [Table 15]

[0348] result Progression-free survival (PFS; Figures 3A and 3C) and overall survival (OS; Figures 3B and 3D) were comparable in ITT patients and in the subsets who were TMB-evaluable for nivolumab monotherapy (Figures 3A and 3B) and nivolumab / ipilimumab combination therapy (Figures 3C and 3D), respectively. ORRs in ITT and TMB-evaluable patients were 11.4% and 11.3% for nivolumab monotherapy and 21.8% and 28.2% for nivolumab / ipilimumab combination therapy, respectively. The TMB distributions for patients receiving nivolumab monotherapy or nivolumab / ipilimumab combination therapy are shown in Figure 4A. When combined (Figure 4B), the distribution of total missense mutations in the SCLC cohort was comparable to that in recent non-small cell lung cancer (NSCLC) clinical trials (Figure 4C).

[0349] The overall response rate (ORR) was higher in TMB-evaluable patients receiving nivolumab / ipilimumab combination therapy (28.2%) than in those receiving nivolumab monotherapy (11.3%) (Figure 5). When stratified by TMB, the greatest benefit was observed in patients with high TMB. Patients with low TMB treated with nivolumab or ipilimumab monotherapy demonstrated ORRs of approximately 4.8% and 22.2%, respectively. Patients with intermediate TMB treated with nivolumab or ipilimumab monotherapy demonstrated ORRs of approximately 6.8% and 16.0%, respectively. Patients with high TMB treated with nivolumab or ipilimumab monotherapy demonstrated ORRs of approximately 21.3% and 46.2%, respectively.

[0350] In general, subjects experiencing a favorable response had a higher number of missense mutations. Subjects receiving nivolumab monotherapy who experienced a complete or partial response (CR) had an average of 325 missense mutations, those experiencing stable disease had an average of 211.5 missense mutations, and those experiencing stable disease had an average of 185.5 missense mutations (Figure 6A). Subjects receiving nivolumab / ipilimumab combination therapy who experienced a complete or partial response (CR) had an average of 266 missense mutations, those experiencing stable disease had an average of 202 missense mutations, and those experiencing stable disease had an average of 156 missense mutations (Figure 6B).

[0351] Furthermore, subjects with high TMB demonstrated increased PFS after treatment with nivolumab monotherapy (Figure 7A) or nivolumab / ipilimumab combination therapy (Figure 7B) compared with subjects with low or intermediate TMB. For nivolumab monotherapy, the mean PFS was approximately 1.3% for low and intermediate TMB subjects and approximately 1.4% for high TMB subjects, with 1-year PFS of 21.2% for high TMB subjects compared with only 3.15% for intermediate TMB subjects (Figure 7A). For nivolumab / ipilimumab combination therapy, the mean PFS was approximately 1.5% for low TMB subjects, 1.3% for intermediate TMB subjects, and approximately 7.8% for high TMB subjects, with 1-year PFS of approximately 30% for high TMB subjects compared with approximately 8.0% and 6.2% for intermediate and low TMB subjects, respectively (Figure 7B).

[0352] Similarly, subjects with high TMB demonstrated increased OS after treatment with nivolumab monotherapy (Figure 8A) or nivolumab / ipilimumab combination therapy (Figure 8B) compared with subjects with low or intermediate TMB. For nivolumab monotherapy, median OS was approximately 3.1% for low TMB subjects, approximately 3.9% for intermediate TMB subjects, and approximately 5.4% for high TMB subjects, and 1-year OS was 35.2% for high TMB subjects compared with approximately 26.0% for intermediate TMB and 22.1% for low TMB subjects (Figure 8A). For nivolumab / ipilimumab combination treatment, the median OS was approximately 3.4% in low TMB subjects, 3.6% in intermediate TMB subjects, and approximately 22% in high TMB subjects, and the 1-year OS was approximately 19.6% and 23.4% in intermediate and low TMB subjects, respectively, compared with approximately 62.4% in high TMB subjects ( Figure 8B ).

[0353] All publications, patents, and patent applications disclosed herein are herein incorporated by reference to the same extent as if each individual publication, patent, and patent application was specifically and individually indicated to be incorporated by reference.

[0354] This application claims the benefit of U.S. Provisional Application Nos. 62 / 572,514 (filed October 15, 2017) and 62 / 650,654 (filed March 30, 2018), which are incorporated herein by reference in their entireties.

Claims

1. A composition comprising an antibody or antigen-binding portion thereof ("anti-PD-1 antibody") that specifically binds to the programmed cell death-1 (PD-1) receptor and inhibits PD-1 activity, for use in treating a subject having a tumor derived from small cell lung cancer (SCLC), wherein the tumor has a tumor mutational burden (TMB) status of high TMB.

2. A composition comprising an antibody or antigen-binding portion thereof that specifically binds to the programmed cell death-1 (PD-1) receptor and inhibits PD-1 activity (an "anti-PD-1 antibody"), in combination with an antibody or antigen-binding portion thereof that specifically binds to CTLA-4 (an "anti-CTLA-4 antibody"), for use in treating a subject having a tumor derived from SCLC, wherein the tumor has a TMB status of high TMB.

3. 3. The composition for use according to claim 1 or 2, wherein the TMB status is determined by sequencing nucleic acid in the tumor and identifying genomic alterations in the sequenced nucleic acid.

4. Genomic changes (i) one or more somatic mutations; (ii) one or more nonsynonymous mutations; (iii) one or more missense mutations; (iv) one or more alterations selected from the group consisting of base pair substitutions, base pair insertions, base pair deletions, copy number alterations (CNAs), gene rearrangements, and any combination thereof; or (v) Any combination of (i) to (iv) 4. The composition for use according to claim 3, comprising:

5. High TMB is at least 210, at least 215, at least 220, at least 221, at least 222, at least 223, at least 224, at least 225, at least 226, at least 227, at least 228, at least 229, at least 230, at least 231, at least 232, at least 233, at least 234, at least 235, at least 236, at least 237, at least 238, at least 239, at least 240, at least 241, at least 242, at least 243, at least 244, at least 245, at least 246, at least 247, at least 248, at least 249, at least 250, at least 255, at least 260, at least 265, at least 270, at least 275, at least 280, at least 285, at least 290, at least 295, at least 300, 5. The composition for use according to any of claims 1 to 4, having a score of at least 305, at least 310, at least 315, at least 320, at least 325, at least 330, at least 335, at least 340, at least 345, at least 350, at least 355, at least 360, at least 365, at least 370, at least 375, at least 380, at least 385, at least 390, at least 395, at least 400, at least 405, at least 410, at least 415, at least 420, at least 425, at least 430, at least 435, at least 440, at least 445, at least 450, at least 455, at least 460, at least 465, at least 470, at least 475, at least 480, at least 485, at least 490, at least 495 or at least 500.

6. 6. The composition for use according to any one of claims 1 to 5, wherein the subject's TMB status is compared to a control TMB value, wherein the subject's TMB status is within the highest quantile of the control TMB values ​​or the subject's TMB status is within the highest tertile of the control TMB values.

7. 7. The composition for use according to any one of claims 1 to 6, wherein the biological sample comprises a tumor tissue biopsy, a liquid biopsy, blood, serum, plasma, exoRNA, circulating tumor cells, ctDNA, cfDNA or any combination thereof.

8. TMB status (i) genome sequencing; (ii) exome sequencing; (iii) genomic profiling or (iv) Any combination of (i) to (iii) The composition for use according to any one of claims 1 to 7, wherein the concentration of hydroxybenzoates is determined by:

9. ゲノムプロファイル、HAR3、HAR2、HARSH102、HAR1 、SATRO、SAT2、SAT1、SAT2、SAT2、 CHRIST、HAR3、KHY1、SHYS、SYS1 1、SHA1K、SHASH2、SHASH、SHASH、SHASH、 WHY、DYS、DYSYS、SYSYS、SYS1 、DY4、DYSYS、DySYS、DY4(SY73). 9)、SYSYS(SYS3)、SYS11、SYS1、SYS3 3、SYS、SYS2、SYS1、SYSYS、SYS 1, HYS, SYS2, SYS, S, S, S, S, S, S, S, S, S CHR3、CH11、CHYCH2、CHY11、CHY 56、SIGN、DYSHY、SYSHY、SYS3、DYS TH、SYSY、SYSY、SYS13、SYSYS、SYSY、1 ROCK1 ROCK1 ROCK1(RO23)、 11030(SYS)、SHSH、SH1S、SHYS、 DAZ、DYSHY、SYS1S1、SHAY(プロモーターのみ)、HARSH、HARSH11、SHARH11、HARSH7 DASH、SHA1、SH1H、SHASH1、SHASH2 、SHE、SIR、SHERS10、SEE10、SHEY124、SEE 4、KHY(HYH1)、DYHY2、SHAY、SHY2 、DAY、DAY3、DAY14、DAY2Y 、KHYK、SHYS2、SHYSHY、SYSHYS3、SHYS1 、KHY1、CHHYH、CHH19、HHH3、HHHHH、N9 88、DYS2、SHOSE、SHOSES14、SHOSE1S HOOD2、SHYS、CHY23、SHYSHY、CHY2(FOUR) 、D1、DYS1、SYS2、SYS1、SYS10、S DAY3、DY2、CHY3、CHY3、CHY2H(CHAS3) 、DY2、DYS、SYS1、SYS22、SYS2、SYS 1. CHRIC1、FEN4、FENSE、CHE20(CHARE2)、9 CHEY22、SYSY12、SYS2、SYS53、SYSY1、 DA274、DYS3、CHS6、DN12、SHAS、9 CHEEK、SHEY1、SHEY2、SHEY1、SHEY、SHEYSYS、DOT1L, FGFR1, HRAS, LMO1, NKX2-1, PREX2, SMAD3, TSC2, ATR, CD79B, EG FR, FGFR2, HSD3B1, LRP1B, NOTCH1, PRKAR1A, SMAD4, TSHR, ATRX, CDC73 , EP300, FGFR3, HSP90AA1, LYN, NOTCH2, PRKCI, SMARCA4, U2AF1, AURKA , CDH1, EPHA3, FGFR4, IDH1, LZTR1, NOTCH3, PRKDC, SMARCB1, VEGFA, AUR KB, CDK12, EPHA5, FH, IDH2, MAGI2, NPM1, PRSS8, SMO, VHL, AXIN1, CDK4 , EPHA7, FLCN, IGF1R, MAP2K1, NRAS, PTCH1, SNCAIP, WISP3, AXL, CDK6, EPHB1, FLT1, IGF2, MAP2K2, NSD1, PTEN, SOCS1, WT1, BAP1, CDK8, ERBB2 , FLT3, IKBKE, MAP2K4, NTRK1, PTPN11, SOX10, XPO1, BARD1, CDKN1A, ERB B3, FLT4, IKZF1, MAP3K1, NTRK2, QKI, SOX2, ZBTB2, BCL2, CDKN1B, ERBB 4, FOXL2, IL7R, MCL1, NTRK3, RAC1, SOX9, ZNF217, BCL2L1, CDKN2A, ERG , FOXP1, INHBA, MDM2, NUP93, RAD50, SPEN, ZNF703, BCL2L2, CDKN2B, ER RFI1, FRS2, INPP4B, MDM4, PAK3, RAD51, SPOP, BCL6, CDKN2C, ESR1, FUBP 10. The composition for use according to claim 8, comprising one or more genes selected from the group consisting of: 1, IRF2, MED12, PALB2, RAF1, SPTA1, BCOR, CEBPA, EZH2, GABRA6, IRF4, MEF2B, PARK2, RANBP2, SRC, BCORL1, CHD2, FAM46C, GATA1, IRS2, MEN1, PAX5, RARA, STAG2, BLM, CHD4, FANCA, GATA2, JAK1, MET, PBRM1, RB1, STAT3, and any combination thereof.

10. (i) SCLC includes small cell carcinoma; (ii) the SCLC includes mixed small cell carcinoma; (iii) the SCLC is relapsed or refractory after at least one prior therapy to treat the tumor; or (iv) any combination of (i) to (iii); A composition for use according to any one of claims 1 to 9.

11. 11. The composition for use of any of claims 1-10, wherein the anti-PD-1 antibody is administered at a body weight-based dose ranging from 0.1 mg / kg to 10.0 mg / kg body weight or at a flat dose of at least about 200 mg, at least about 220 mg, at least about 240 mg, at least about 260 mg, at least about 280 mg, at least about 300 mg, at least about 320 mg, at least about 340 mg, at least about 360 mg, at least about 380 mg, at least about 400 mg, at least about 420 mg, at least about 440 mg, at least about 460 mg, at least about 480 mg, at least about 500 mg, or at least about 550 mg every two, three, or four weeks.

12. Anti-PD-1 antibody (i) a weight-based dose of 3 mg / kg body weight every 2 weeks; (ii) a weight-based dose of 5 mg / kg body weight every 3 weeks; (iii) a weight-based dose of 10 mg / kg body weight every 3 weeks; (iv) a fixed dose of about 240 mg every two weeks; or (v) a fixed dose of approximately 480 mg every 4 weeks The composition for use according to any one of claims 1 to 11, which is administered to

13. 13. The composition for use according to any of claims 2 to 12, wherein the anti-CTLA-4 antibody is administered at a dose ranging from at least about 0.1 mg / kg to at least about 10.0 mg / kg of body weight about every 1, 2, 3, or 4 weeks.

14. Genomic profiles are available in FOUNDATIONONE® CDX TM 9. The composition for use according to claim 8, comprising:

15. 15. The composition for use according to any of claims 1 to 14, wherein the tumor has at least about 10 mutated TMBs per megabase of the sequenced genome.

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